Massimo, I totally agree! I was just about to voice the same opinions. I too am surprised (appalled!) to find such narrow-mindness and xenophobia among supposedly open-minded "scientists". These folks have been looking down their very narrow little tubes far too long and have severe micro-tunnel vision.
} Introduce oil? Don't we have enough trouble with section contamination? } If you chealate the two chemicals properly, you will not have to worry } about carbon dioxide. Keep your solution in a plastic syringe! Drive all } the air out of it. It will keep for years in the refrigerator. Put a } clean 0.22 micrometer filter on the syringe before use. Wet the filter } with the solution before forcing a drop out which is to be used for a } grid. } } Hildy
I switched from Reynold's recipe to Haniachi's version (Hanichi et al. 1986. A stable lead modification of Sato's method. J. Electron. Microsc. 35: 304-306) several years ago and have been quite pleased with the results. Although I could get Reynold's to work for me, I was never 100% successful and my student's grids were rarely precipitate free. Now, we use a single 30-60 sec stain with the calcined lead and don't even bother with a uranium secondary contrast (radioactive wastes have become a thorny issue here). I work mostly with Spurr's embedded plant material but we also do Epon embedded animal tissues for EM class; both have nice contrast.
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
I am not going to Cancun, but rather wish I was. However, I was just asked by a friend who has a son leaving for Mexico, what is needed by way of proof of identity for travel to Mexico? I know that neither passport or visa is necessary for travel to Canada. Is the same true of our southern neighbor?
At 02:52 PM 7/31/98 +1000, you wrote: } Dry oxygen maybe a lesser consideration, however, when the filament chamber } is to be vented, it's even more important to wait until the filament has } cooled. Exposure of a hot filament or LaB6 cathode to N2 is not a good idea, } exposure to O2 is worse. } Cheers } Jim Darley } ProSciTech Microscopy PLUS
Hmmm. We vent our Hitachi with W filament with He (because of less scattering effect compared to N2). Should we be concerned about any reaction between the hot filament and the He? I would presume none. How about thermal shock effects? Anybody care to comment?
No offense intended, but this discussion is getting way too anal. I'm not sure what 'scattering' effects you believe helium will improve. If you are referring to electron scattering during operation, helium is probably harder for normal vacuum systems to pump out then nitrogen, so the gain you are seeking more than likely isn't there. I know of no practical difference between venting with any dry gas.
Conductive heat dissipation is always much faster than convective. In the two or three seconds it takes you switch the accelerating voltage off and to hit the vent button, the majority of the heat built up in the small mass of a tungsten filament will probably be conducted off through the filament posts and electrical contacts in the gun. As the backfill gas works its way into the gun, the remainder of the heat will be removed rather slowly since the leak provided by a normal vent valve is rather small, taking 30 seconds or more to completely vent.
I have never seen or heard of anything other than very loose anecdotal evidence for increases in cathode life dependent on venting methods. At the most, I'll accept customers turning the filament drive down before turning off the accelerating voltage and venting the instrument. Anything more than that is simply unnecessary. It is far more important to be cognizant of small vacumm leaks, particularily in areas that could bleed into the gun area. Air leaks in the gun during operation are extremely damaging.
} At 02:52 PM 7/31/98 +1000, you wrote: } } Dry oxygen maybe a lesser consideration, however, when the filament } } chamber is to be vented, it's even more important to wait until the } } filament has cooled. Exposure of a hot filament or LaB6 cathode to } } N2 is not a good idea, exposure to O2 is worse. Cheers Jim Darley } } ProSciTech Microscopy PLUS } } Hmmm. We vent our Hitachi with W filament with He (because of less } scattering effect compared to N2). Should we be concerned about any } reaction between the hot filament and the He? I would presume none. } How about thermal shock effects? Anybody care to comment? Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Note: Entries for the 14th ICEM Photomicrograph Exhibit will be accepted through August 15th.
ICEM 14 - Cancun PHOTOMICROGRAPHS ON DISPLAY
There will be a Photomicrograph Exhibit held in conjunction with the 14th International Congress on Electron Microscopy in Cancun this August 31 - September 4. If you are attending the meeting, please consider bringing up to 3 micrographs for inclusion in a Photomicrograph Exhibit. From this exhibit, images will be selected for inclusion in a post conference publication. See details below and if you have questions contact either the meeting headquarters in Mexico City (info below) or myself.
Photomicrograph Exhibition
The 14th ICEM will include a micrograph exhibition. The organizers' intentions are to provide a showcase for outstanding micrographs displaying a combination of art and science.
The exhibit is open to all forms of microscopic imaging. A panel of judges will select entries based on artistic merit for inclusion in a post-congress publication of images. Submitted micrographs much be accompanied by a brief description, not only of their scientific content, but also of their technical aspects. If images have been digitally processed or altered, the digital processing should be described as well. All submitted micrographs will be displayed although entries not regarded by the judges as artistically significant will be excluded from the publication of images. Please read the following rules carefully:
Only registered attendees at the 14th ICEM are eligible to submit micrographs.
Any individual may submit up to three micrographs.
Entries must be 27.5 cm x 35.0 cm, may be mounted vertically or horizontally, and must be affixed to a stiff lightweight support, such as 10 mm foam board. Micrographs may either be flush mounted or have borders so long as the overall dimensions of the entry are 27.5 cm x 35.0 cm.
Entries must be brought to the meeting and mounted on the display boards by the entrant or his/her delegate. Velcro will be provided. Entries must be mounted between 12:00 and 17:00 on Monday, August 31st and those micrographs not selected for publication must be removed between 12:00 and 15:00 on Friday, September 4th. Micrographs remaining on the display boards after then will be discarded. Micrographs selected for publication will not be returned to the entrant and will become the property of the 14th ICEM.
Selected micrographs will be incorporated into a post-congress publication of images. Those selected for inclusion will be announced during the Thursday evening reception.
To enter: Send your name, address, telephone and fax numbers, and email address plus a description of 200 words or less for each entered micrograph (maximum of 3) to: Secretariat 14th International Congress on Electron Microscopy Amsterdam 46-202 Col. Hipodromo Condesa C.P. 06100, Mexico, D.F. MEXICO Phone: (525) 553-4507; Fax (525) 553-4500 email: icem-at-icem.inin.mx
Entry information must be received by August 15, 1998. Entries will be acknowledged promptly. Do not send micrographs, you must bring them or have them brought to the Meeting.
Meeting Office staff will print your description(s) in standard form and prepare them to be mounted along with your micrograph(s) at the Meeting.
___________________________________________________________________________ Barbara Reine, Botany Dept. Box 351330 Univ. of Washington, Seattle, WA 98195-1330 e-mail: reine-at-u.washington.edu; Phone: (206) 543-1955; Fax: (206) 543-3262 ____________________________________________________________________________
I should have mentioned that our Hitachi is a 2460N low vacuum SEM. During operation we He instead of air or N2 in order to reduce scattering effects at the 40 Pa of atmosphere that we maintain in order to cancel charging on our specimens. There was no intent of trying to reduce scatter by residual gas at 10-5 torr levels since He might have displaced heavier molecules.
I was intrigued that a side benefit of our He use might be increased filament life since the hot filament would not be exposed to N2 with its detrimental effects. A coworker likes to wait until the filament cools for a bit before venting the chamber (the gun chamber always gets vented with the specimen chamber). However, I had been supposing that thermal issues might be insignificant as you appear to corroborate. Since we just had a W filament go 441 hours even with sample changes about every 2 hours, I suppose that the Helium might be helping us some. However the benefits are probably not worth someone switching over from N2 to He unless they too have a "leaky" SEM.
Warren Straszheim
At 03:32 AM 8/2/98 -0600, Allen R. Sampson wrote: } No offense intended, but this discussion is getting way too anal. } I'm not sure what 'scattering' effects you believe helium will } improve. If you are referring to electron scattering during } operation, helium is probably harder for normal vacuum systems to } pump out then nitrogen, so the gain you are seeking more than likely } isn't there. I know of no practical difference between venting with } any dry gas. } } Conductive heat dissipation is always much faster than } convective. In the two or three seconds it takes you switch the } accelerating voltage off and to hit the vent button, the majority of } the heat built up in the small mass of a tungsten filament will } probably be conducted off through the filament posts and electrical } contacts in the gun. As the backfill gas works its way into the } gun, the remainder of the heat will be removed rather slowly since } the leak provided by a normal vent valve is rather small, taking 30 } seconds or more to completely vent. } } I have never seen or heard of anything other than very loose } anecdotal evidence for increases in cathode life dependent on venting } methods. At the most, I'll accept customers turning the filament } drive down before turning off the accelerating voltage and venting } the instrument. Anything more than that is simply unnecessary. It } is far more important to be cognizant of small vacumm leaks, } particularily in areas that could bleed into the gun area. Air leaks } in the gun during operation are extremely damaging. } } } At 02:52 PM 7/31/98 +1000, you wrote: } } } Dry oxygen maybe a lesser consideration, however, when the filament } } } chamber is to be vented, it's even more important to wait until the } } } filament has cooled. Exposure of a hot filament or LaB6 cathode to } } } N2 is not a good idea, exposure to O2 is worse. Cheers Jim Darley } } } ProSciTech Microscopy PLUS } } } } Hmmm. We vent our Hitachi with W filament with He (because of less } } scattering effect compared to N2). Should we be concerned about any } } reaction between the hot filament and the He? I would presume none. } } How about thermal shock effects? Anybody care to comment? } Allen R. Sampson } Advanced Research Systems } 317 North 4th. Street } St. Charles, IL 60174 } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net } WWW: http://www.mcs.net/~ars } Analytical instrument maintenance services }
} } Put a clean 0.22 micrometer filter on the syringe before use. } } Wet the filter with the solution before forcing a drop out } } which is to be used for a grid.
We have found that using a filter is not necessary. If the solution has been made-up and stored properly it does not require filtration.
} Hildy: I started dispensing lead citrate from a syringe recently, and find } it works pretty well. However, would you clarify/elaborate the part 'Wet } the filter...' ?
As I reported earlier, we store the made-up Reynolds in approx. 3 ml aliquots in Eppendorff tubes. We dispense by taking up the stain into a new disposable Pasteur pipette from about 4 - 5mm above the bottom of the tube and then discard the first drop. This minimizes the possibility that any contamination from crystals, dirt, etc, which may have accumulated at the bottom of the tube, on the surface of the stain or in the pipette will end up in the staining drop. Once the Eppendorff tube has been opened it is not re-used - it is discared with any left-over stain.
This uncomplicated method has produced almost totally contamination-free stain for many years, even in the hands of the most novice workers.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
Rick Felten wrote on 07/31/98 11:39 AM: } I would like to determine the thickness of Au and Ag on a copper substrate. } The copper is hundreds of microns thick; the Ag is a few thousand angstroms } and the Au is a few hundred angstroms. When the thickness of Ag is } required a Ag on Copper sample could be submitted (no Au). We have the } ability to make up varying (but unknown) thickness of Ag and Au and could } try to plate and cross section at a high angle or digest the deposit and } analyze with AA to get thickness standards. Or is it better to use pure Au } and pure Ag standards and let theory and computer software do the rest. } } Thanks } Ric
Hi Ric,
I once did some measurements of layer thickness of Au on Ho by the Bishop-Poole method (H. E. Bishop and D. M. Poole, J. Phys. D: Appl. Phys., vol. 6 (1973) 1142) with the use of a pure Au standard. The results were in good agreement with the thicknesses expected from the deposition parameters, so I think you can safely use pure standards and "let theory and computer software do the rest".
Best regards, Jorgen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
My old Link AN10000 system has a software package called TFOS (thin films on substrate) which does this calculation. I haven't used it for years but I seem to remember that it worked OK.
Regards
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
} } Rick Felten wrote on 07/31/98 11:39 AM: } } I would like to determine the thickness of Au and Ag on a } copper substrate. } } The copper is hundreds of microns thick; the Ag is a few } thousand angstroms } } and the Au is a few hundred angstroms. When the thickness of Ag is } } required a Ag on Copper sample could be submitted (no Au). We have the } } ability to make up varying (but unknown) thickness of Ag and Au } and could } } try to plate and cross section at a high angle or digest the deposit and } } analyze with AA to get thickness standards. Or is it better to } use pure Au } } and pure Ag standards and let theory and computer software do the rest. } } } } Thanks } } Ric }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
REGARDING in search for FEG-STEM in Europe
This message was forwarded to me by Bruce Davidson at = davidson-at-fisica.cib.na.cnr.it. Please respond directly to him.
In search of a cold-stage FEG-STEM in Europe
My name is Bruce Davidson from the Istituto di Cibernetica -CNR in = Naples, Italy. I'm searching for a FEG-STEM with cold-stage in Europe in = conjunction with a project to scribe Josephson junctions in YBCO thin films using a ~200 keV FEG source at ~150 K.
The idea for these junctions is to create a narrow region of oxygen displacement in a link patterned in a YBCO film by scanning a ~1 nm beam of electrons across the link once. The amount of oxygen = displacement determines the Tc reduction in the irradiated region, which cannot be = wider than a few nanometers in order to observe Josephson coupling across it. These junctions are some of the best-behaving, most uniform junctions = made to date in the high-temperature superconductors. We have fabricated many of these junctions in the past few years (the subject of my PhD thesis at = the Univ. of Wisconsin, August '97).
I am currently in Italy on a NSF-NATO Post-doc fellowship to expand the study of this technique. I would like to perform the irradiation of the = films using a STEM here in Europe rather than one in the US, if a suitable microscope can be found. The essential capabilities are:
1) FEG electron source (~0.5 nA current, ~1 nm FWHM spot -at- 200 keV). 2) scanning capabilities, with computer control of the scan coils. 3) sample cold stage, to maintain the sample at ~150 K during = irradiation.
Any information on suitable microscopes in Europe would be greatly appreciated.
A colleague is having problems getting good fixation of Xenopus neurons grown in culture, the membranes are shot by both SEM and TEM. She cannot add calcium to the fix due to certain experimental conditions. Any help would be greatly appreciated.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
If you have a diffusion pump or turbopumped SEM, the pump speed for different gas species goes as the molecular weight of the molecule. As a result, the pump speed for He compared to N2 are much lower. I could look it up, but I think that it is about an order of magnitude different. Another point is that, because the sensitivity of ion gauges is about 0.25 for He, the gauge pressure and the true pressure will be different. With a sensitivity of 0.25 for an ion gauge, the true pressure is four times the value displayed. I'm not sure what the values are for Penning gauges, but they are probably in the same ball park.
If your vacuum system has ion pumps, DO NOT backfill with an inert gas, especially He! Even with the special ion pumps that bury the inert gases into the plates, they still "burp" the inert gas. Helium pump speeds are extremely low.
One interesting aspect about your use of He around a hot filament is that it does have a higher thermal conductivity than N2, so I guess that it would cool the filament faster.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I should have mentioned that our Hitachi is a 2460N low vacuum SEM. During operation we He instead of air or N2 in order to reduce scattering effects at the 40 Pa of atmosphere that we maintain in order to cancel charging on our specimens. There was no intent of trying to reduce scatter by residual gas at 10-5 torr levels since He might have displaced heavier molecules.
I was intrigued that a side benefit of our He use might be increased filament life since the hot filament would not be exposed to N2 with its detrimental effects. A coworker likes to wait until the filament cools for a bit before venting the chamber (the gun chamber always gets vented with the specimen chamber). However, I had been supposing that thermal issues might be insignificant as you appear to corroborate. Since we just had a W filament go 441 hours even with sample changes about every 2 hours, I suppose that the Helium might be helping us some. However the benefits are probably not worth someone switching over from N2 to He unless they too have a "leaky" SEM.
Warren Straszheim
At 03:32 AM 8/2/98 -0600, Allen R. Sampson wrote: } No offense intended, but this discussion is getting way too anal. } I'm not sure what 'scattering' effects you believe helium will } improve. If you are referring to electron scattering during } operation, helium is probably harder for normal vacuum systems to } pump out then nitrogen, so the gain you are seeking more than likely } isn't there. I know of no practical difference between venting with } any dry gas. } } Conductive heat dissipation is always much faster than } convective. In the two or three seconds it takes you switch the } accelerating voltage off and to hit the vent button, the majority of } the heat built up in the small mass of a tungsten filament will } probably be conducted off through the filament posts and electrical } contacts in the gun. As the backfill gas works its way into the } gun, the remainder of the heat will be removed rather slowly since } the leak provided by a normal vent valve is rather small, taking 30 } seconds or more to completely vent. } } I have never seen or heard of anything other than very loose } anecdotal evidence for increases in cathode life dependent on venting } methods. At the most, I'll accept customers turning the filament } drive down before turning off the accelerating voltage and venting } the instrument. Anything more than that is simply unnecessary. It } is far more important to be cognizant of small vacumm leaks, } particularily in areas that could bleed into the gun area. Air leaks } in the gun during operation are extremely damaging. } } } At 02:52 PM 7/31/98 +1000, you wrote: } } } Dry oxygen maybe a lesser consideration, however, when the filament } } } chamber is to be vented, it's even more important to wait until the } } } filament has cooled. Exposure of a hot filament or LaB6 cathode to } } } N2 is not a good idea, exposure to O2 is worse. Cheers Jim Darley } } } ProSciTech Microscopy PLUS } } } } Hmmm. We vent our Hitachi with W filament with He (because of less } } scattering effect compared to N2). Should we be concerned about any } } reaction between the hot filament and the He? I would presume none. } } How about thermal shock effects? Anybody care to comment? } Allen R. Sampson } Advanced Research Systems } 317 North 4th. Street } St. Charles, IL 60174 } PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net } WWW: http://www.mcs.net/~ars } Analytical instrument maintenance services }
For US citizens going to Mexico, you can use a passport, an out of date passport, birth certificate, or a certified copy of your birth certificate. -Scott ----------
I am not going to Cancun, but rather wish I was. However, I was just asked by a friend who has a son leaving for Mexico, what is needed by way of proof of identity for travel to Mexico? I know that neither passport or visa is necessary for travel to Canada. Is the same true of our southern neighbor?
Thanks to all who replied about necessary travel documentation. It looks like passports are still not officially required but would be advisable. I know I got a few mean looks coming back from Canada without one a couple years ago. As long as it doesn't get any worse than the mean looks, my friend should be fine.
The reason you are getting breakage is due to the shrinkage that processing and CPD in particular causes--up to 50% in some tissues. Since these are adherent cells, those neurites which are firmly adhered will probably show some form of breakage/damage. The only improvements I have obtained (and not consistently, I admit) has been using products such as Peldri or one of the hexamethyldisilazane products. This method is less consistent than CPD, but can yield better preservation of morphology of fragile samples. Contact any of the EM supply houses (EMS, SPI, Pella, Energy Beam Science, Fullam, Ladd, Polysciences) and ask for their technical fact sheets. There is also a fair amount of published methods literature--primarily from the 1980s. Hope this helps.
Roger Moretz, Ph.D. Toxicology
The comments are solely the opinions of the author. I have no vested interest in any of the products or the suppliers.
} -----Original Message----- } From: Sandy Perkins [SMTP:skperkin-at-vt.edu] } Sent: Friday, July 31, 1998 10:08 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM-neuroblastoma cells } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } Hi- } } We are trying to refine our procedure for the preparation of } differentiated } neuroblastoma cells for SEM. The cells are grown on coverslips and we } have } used the following processing schedule: } } gentle phosphate buffer wash (0.1 M sodium phosphate, pH 7.4) } primary fixation in 2.5% glut/1% paraformaldehyde-1 hr } buffer washes } post-fixation in 1% osmium tetroxide-1 hr } ethanol dehydration } CPD } } The cells look OK, but a number of the neurite extensions are broken. } Has } anyone had any experience processing these cells for SEM? Do we need } a } "gentler" processing method? Any help would be greatly appreciated. } Thank } you very much. } } Sandy Perkins } } Laboratory for Neurotoxicity Studies } VA Tech }
I would be curious if anyone has any real data on filament life as affected by a cooling off period before chamber venting. Like many, I was trained to always cool a filament for several minutes before admitting air, but if it's really not necessary I'd be happy to stop. Sure would speed specimen turn-around times when we're running a bunch through.
Another question: do you really find it necessary to use 40 pa to eliminate charging on your 2460? Using both a 2460 and a 3200, I have almost always been able to eliminate charging with pressures of 1-5 pa, which should give substantially less scattering than 40 pa. Just wondering.
Finally---filament life of more than 400 hours is amazing!! And under variable pressure with frequent specimen changes! I'd love to know the secret to that trick. Maybe He is the wonder gas we've all been waiting for....
Regards,
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Hi: I operate a sem-edxra system, metallurgical microscope, LM and use Image Pro + and supporting equipment for Hallmark Cards Inc. (we also have an FTIR microscope). Over the years (20) I have been asked to do many unusual projects . One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days (old equipment and user). I have talked to a couple of users of and one supplier of microtomes and they seem to think I need one (surprise) . I don't know anything about microtomes what type (rotary / sliding) and what blades or what mounting media would be most useful for us. There seem to be only one supplier of microtomes (Leica) are there any others? Does anyone have any recommendations or experience with this type of materials? Of course I will have to come up with justifications to buy one and any help would be appreciated. Thanks Terry Ellis email-tellis2-at-hallmark.com 816-545-6573 mail drop 359 2501 McGee, box 419580 K.C. MO 64141-6580
Randy writes ... } } } I would be curious if anyone has any real data on filament } life as affected by a cooling off period before chamber venting. } Like many, I was trained to always cool a filament ...
I'm not sure "real data" is needed. Tungsten's affinity for oxidation at elevated temperatures is well known. I believe most of us who don't wait for a cool down period are venting with dry nitrogen or some other inert gas.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Hi: I operate a sem-edxra system, metallurgical microscope, LM and use Image Pro + and supporting equipment for Hallmark Cards Inc. (we also have an FTIR microscope). Over the years (20) I have been asked to do many unusual projects . One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days (old equipment and user). I have talked to a couple of users of and one supplier of microtomes and they seem to think I need one (surprise) . I don't know anything about microtomes what type (rotary / sliding) and what blades or what mounting media would be most useful for us. There seem to be only one supplier of microtomes (Leica) are there any others? Does anyone have any recommendations or experience with this type of materials? Of course I will have to come up with justifications to buy one and any help would be appreciated. Thanks Terry Ellis email-tellis2-at-hallmark.com 816-545-6573 mail drop 359 2501 McGee, box 419580 K.C. MO 64141-6580
Hi: I operate a sem-edxra system, metallurgical microscope, LM and use Image Pro + and supporting equipment for Hallmark Cards Inc. (we also have an FTIR microscope). Over the years (20) I have been asked to do many unusual projects . One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days (old equipment and user). I have talked to a couple of users of and one supplier of microtomes and they seem to think I need one (surprise) . I don't know anything about microtomes what type (rotary / sliding) and what blades or what mounting media would be most useful for us. There seem to be only one supplier of microtomes (Leica) are there any others? Does anyone have any recommendations or experience with this type of materials? Of course I will have to come up with justifications to buy one and any help would be appreciated. Thanks Terry Ellis email-tellis2-at-hallmark.com 816-545-6573 mail drop 359 2501 McGee, box 419580 K.C. MO 64141-6580
My apologies for not being more specific in my first post. The question should have been "Does anyone have a recommendation for a good TEM/SEM fixative for Xenopus neurons grown in culture?" My colleague has had very poor TEM results with 2.5% glut in 0.1M cacodylate with 0.1M sucrose, there are no membranes visible. Cell membranes, mitochondrial membranes, organelle membranes, all gone/invisible. Yes, post-fixation with osmium was used. Overall ultrastructure is also poor. She has also had very poor SEM results with the same fix with the addition of 4% paraformaldehyed, plasma membrane had large holes. When I suggested adding calcium to the fix I was told that the experimental protocol prohibited this. Any help will be appreciated!
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
At 11:54 AM 8/3/98 -0600, Randy Tindall wrote: } I would be curious if anyone has any real data on filament life as affected } by a cooling off period before chamber venting. Like many, I was trained } to always cool a filament for several minutes before admitting air, but if } it's really not necessary I'd be happy to stop. Sure would speed specimen } turn-around times when we're running a bunch through.
One operator likes to cool the filament for a while to get longer life. But this filament has had a number of quick cool downs anyway. If it is a reaction with nitrogen, then maybe we can vent the He to it right away. Then again, we ran the scope in automatic mode with a beam shut off at the end of the run. It might have had plenty of time to cool for most exchanges.
} Another question: do you really find it necessary to use 40 pa to eliminate } charging on your 2460? Using both a 2460 and a 3200, I have almost always } been able to eliminate charging with pressures of 1-5 pa, which should give } substantially less scattering than 40 pa. Just wondering.
We have tried reducing the pressure and have run into charging. But we run a lot of beam current to our concrete samples, around 1-2 nA by my guess. Maybe the lower pressures work for lower beam current.
} Finally---filament life of more than 400 hours is amazing!! And under } variable pressure with frequent specimen changes! I'd love to know the } secret to that trick. Maybe He is the wonder gas we've all been waiting } for....
I just hope that we can replicate the performance. Our normal life has been in the 100 hour range.
I have been asked to organize a session on precision specimen prep for the 1999 MSA/MAS meeting in Portland. Here is the descriptive paragraph for the Call for Abstracts:
======
Precision Specimen Preparation
In the physical sciences we see an increasing need to prepare TEM specimens of very small pre-selected locations in a variety of different types of samples. The main driving force for high spatial resolution specimen preparation is the semiconductor industry, where the size of the target locations to be prepared have diminished to a point where they cannot be viewed in a visible light microscope. Precision preparation of the impact point of a projectile into armor or an analysis of the precise initiation point of a micro-crack are other examples. This symposium seeks to review the state of the art of precision specimen preparation and to explore the prospects for improved spatial resolution in the near future.
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As I've been correctly accused of having a bias towards tripod polishing, I am looking for suggestions (self-suggestions are fine!) for people to ask to be Invited Speakers in this symposium that would bring a broader perspective to the topic. I would be especially interested in someone to address non-semiconductor examples. I could use a symposium co-chair too, if there is one out there.
Now that MSA's middle school microscopy manual is published, Project MICRO (Microscopy In Curriculum - Research Outreach) is going into high gear. Several MSA local societies already have outreach programs, or are about to begin one (Arizona, Minnesota, North Carolina, Oklahoma,). So Nestor and I plan to expand the MICRO web page (yes, that means that the wonderful microscopy quotes that you folks sent last spring will be posted soon). Many of you have been visiting schools for a long time, and you've developed good materiel. One resource that we want to offer is a collection of successful classroom-tested exercises that can be used to extend the experiences provided by "Microscopic Explorations". Grade level? Elementary & middle school. Topic? Anything that works well. Reward? Our thanks, and the opportunity to share the results of your hard work. Please respond directly, since your text may be lengthy; I'll organize and post.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
As you all know the cathode assembly gets up to a pretty high temperature=
too! If we vent with "dirty" air we cause contamination to deposit on th= e hot cathode. I agree the filament cools very quickly but we all know the=
cathode does not?
Waiting a few minutes after getting the READY indication before switching=
on the filament and a few minutes after switching off the filament, prior=
to letting in AIR, the result a difference in filament life and cathode contamination. =
The first point is that the READY indication means the vacuum is only jus= t good enough to use. Wait a few more minutes and the improvement in vacuu= m will help give a better filament life and keep the column clean longer. =
Put a beam down a column with a poor vacuum and you put up the contamination! From the other direction allowing the cathode to cool pri= or to letting in air will keep this cleaner too!
Just the same when you clean a column, it makes sense to clean the column=
on a Friday afternoon, pump and wait over the weekend before putting a beam down the column. Clean a column and then pass a beam through it means that most of the vapours from the cleaning media have just deposite= d on your clean components!
Steve Chapman Senior Consultant E.M. Protrain for courses and consultancy in electron microscopy world wide Tel & Fax 44 (0)1844 353161 WWW http://ourworld.compuserve.com/homepages/protrain
Terry Ellis wrote: =============================================== ..........One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days ...... ================================================ We have been doing this kind of sample for some number of years and the microtome manufactures are correct, you really can only do this kind of study properly via the use of an ultramicrotome (and diamond knives, glass would be a waste of time). Because the acrylic binders, both in the ink and the paper substrate, have low Tg's, the sectioning in general has to be done cryo. The views of the structure by examining the sections by TEM are so far superior to the best we have been able to obtain by SEM that now we rarely even make the SEM attempt. Questions of ink and pigment penetration you can not come close to in an SEM anyhow.
There is a "trick" that you have to employ to keep the ink from literally dissolving in (or being swollen by) the embedding resin and that is to sputter coat a layer of platinum (we prefer Pt for this purpose over Au because it is less likely to smear during sectioning) onto the surface of interest and THEN do the embedding. Since paper has some amount of porosity , to be on the safe side, we always do a vacuum embedding. If the paper has been in a humid environment, it might be wise to just "pump on it" in a vacuum evaporator over night to pull of some of that unwanted moisture.
We have consistently gotten our best results with our own SPI-Pon™ 812 resin system (but I suspect that at least some of the "Epon replacements" from others would work equally well). With a little bit of practice, you can get the resin to cure in a way that there is virtually no interaction with the sample underneath the Pt (or Au) passivation layer. This layer has the added benefit of making very clear that you are seeing the entire ink layer cross-section and not seeing an artifact because part of it broke away during the sectioning.
The other point has to do with the selection of the diamond knife. There are two considerations, and this part of my posting might be "controversial" : a) Use only "materials science" diamond knives for this kind of work, you will save a lot of money if you do that, and b) follow our (on the website) advice about knife angle, use a 45 deg. angle and not the 55 deg. angle sometimes touted as being "right" for "materials science" work. At least for these kinds of samples, if you have any kind of serious compression effects present, the layers will quite readily split apart, so you want to minimize compression effects by using a 45 deg. angle knife. Will the "sharper" knife edge "wear out faster"? Probably, but not as much as you might think since when compared to a 55 deg. knife, remember that in order to get good sections, you have to make many more sections just to get to that point. And that puts all kinds of additional wear and tear on the knife edge! So the economics are not exactly what common sense might suggest!
The last point has to do with the data presentation. Again, it has been our experience that the deposition of ink is inherently not necessarily all that homogeneous of a process and therefore, we take the micrographs as a montage along the surface (ink) layers. The objective is to show the entire "forest" and not just individual trees. It is the appearance of the forest that more often than not, shows the differences between samples that behave differently. If you are not doing these kinds of samples as montages, you could very easily "miss" what would otherwise be there to be seen.
Disclaimer: SPI Supplies offers the embedding resins and diamond knives to do this kind of work and if this all sounds too complicated to gear up to do it yourself, the laboratory services part of our company would be happy to talk to you about doing it for you.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
In a message dated 98-08-03 15:36:02 EDT, tellis2-at-hallmark.com writes:
{ { One ongoing type of project I haven't been able to consistently do is to cross-section printed paper and measure the clay /clear top coat coating thickness' and determine the ink penetration of various inks. I have been able to use metallurgical procedures to obtain the clay coating thickness' but LR white hard mounting media makes most of the inks run plus the polishing takes 2 to 4 days (old equipment and user). I have talked to a couple of users of and one supplier of microtomes and they seem to think I need one (surprise) . I don't know anything about microtomes what type (rotary / sliding) and what blades or what mounting media would be most useful for us. There seem to be only one supplier of microtomes (Leica) } }
Hi Terry,
I'll sidestep the issue of manufacturers of microtomes, since I'm sure you will hear from all of us who manufacture and sell them. And I like to avoid flames from the SysOp whenever possible...
Actually, I used to do some similar work on paper products in one of my previous lives, and I can enthusiastically recommend cryomicrotomy as the way to go. You can freeze the specimen directly in one of several aqueous or non- aqueous embedding media and section it with either glass, diamond or tungsten carbide knives.
I would recommend a rotary microtome with a cryosectioning kit. The cryo kit is a box that is cooled with liquid nitrogen and has temperature regulation and heaters built in for both the specimen and the knife. The cryobox fits on the microtome in place of the regular knife holder.
If you have a cryostage on the SEM you can transfer the sectioned ("polished") material directly to the scope for evaluation. If not, you can melt the sectioned material out of the embedding media, rinse quickly, dry, then mount and examine in the SEM.
Of course you can also collect the sections and mount them for FTIR.
The main advantages of cryotechniques are bypassing all of the solvents, plastics, abrasives/polishing, etc. that are normally used in specimen prep, and the quick turnaround time. From start to finish, it takes maybe 10 minutes after the cryosectioning device is cooled down.
And yes, Leica makes such equipment. So do a couple of others. I'm sure they will also respond.
Best regards, Bob ***************************************** Robert (Bob) Chiovetti rchiovetti-at-aol.com E. Licht Company / 1-800-865-4248 Colorado/Utah/Wyoming/Arizona/ New Mexico/West Texas U.S.A. Representing Leica Since 1967 *****************************************
Forgive me, Nestor, if this violates your directive about Cancun.
But there have been some rather major fare reductions to Cancun and if you call your airline, you might quality for a nice refund of the difference. That has just saved the two of us a cool $140 on two tickets. At least it worked that way on United.
I expect that filament life has nothing to do with the use of any inert gas. In the 1950th (no, I was not in labs then), a paper was published The life of filaments. My long-term memory recall advises that short of physical violence, filament life is determined by vacuum, if it's under 5x10-5 torr and by emission if it's above 15microamps. It is assumed that throughout it's life the filament is correctly saturated. I recall that there was a curve (or maybe just a number of data points) showing filament "death" at various vacua and emissions. In modern instruments, emission is most frequently the limiting factor. Reaching a filament life of 500 hours is no trouble in microprobes at say 10 to 15 microamps, it's a little less for a TEM, which commonly run at 30 microamps. Tell me when a SEM operating at 80 or more microamps reaches 500 hours filament time; then it will be time to rewrite that 1950ish paper. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Tuesday, 4 August 1998 3:55, Randy Tindall [SMTP:rtindell-at-NMSU.Edu] wrote: . . . . . . } Another question: do you really find it necessary to use } 40 pa to eliminate } charging on your 2460? Using both a 2460 and a 3200, I } have almost always } been able to eliminate charging with pressures of 1-5 pa, } which should give } substantially less scattering than 40 pa. Just wondering. } } } Finally---filament life of more than 400 hours is } amazing!! And under } variable pressure with frequent specimen changes! I'd } love to know the } secret to that trick. Maybe He is the wonder gas we've } all been waiting } for.... } } Regards, } } Randy } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } } rtindell-at-nmsu (work) } nrtindall-at-zianet.com (home)
your travel agent or the airline will give you a tourist card (actually a flimsy set of thin papers). There is no charge. Part is collected or stamped when you enter, and the remainder is surrended when you leave. The other docs are useful.
I have been noticing a black precipitate on my grids. At high magnification, the precipitate looks like large hexagon chrystals. When the electron beam hits the precipitate, it tears the tissue. Any good hints as to what could be my problem would be of great help.
Moreover, would you happen to know of any jobsites on the net devoted to microscopy?
Thank you for your time,
Maria L. Sunio
P.S. Please email me directly to sunio-at-utsw.swmed.edu
This message is in MIME format. The first part should be readable text, while the remaining parts are likely unreadable without MIME-aware tools. Send mail to mime-at-docserver.cac.washington.edu for more info.
We have great pleasure to inform you about our conference "COLOR '99" organised in Krakow under the auspices of Foundry Research Institute, Krakow, Poland.
Below we send to you the first announcement of this conference in the file Word 7.0 "First.doc".
Best regards.
Members of Local Conference Committee:
Janina Radzikowska and Krzysztof Jan Huebner
================================================
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
} [skip all the stuff I agree with] Clean a column and then pass a beam } through it means that most of the vapours from the cleaning media have } just deposited on your clean components! } Even worse. The radiolysis products--ions, free radicals, etc.-- will deposit on and possibly interact with the column. Yours, Bill Tivol
We had success using 1% glutaraldehyde/0.25% acrolein in 10mM MgCl2, 50mM PIPES, pH 6.5. See Luther et al, J. Neurocytol 25, 417-427, 1996.
Steve Samuelsson ______________________________ Reply Separator _________________________________
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Dear List:
A colleague is having problems getting good fixation of Xenopus neurons grown in culture, the membranes are shot by both SEM and TEM. She cannot add calcium to the fix due to certain experimental conditions. Any help would be greatly appreciated.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
by imo28.mx.aol.com (IMOv14_b1.1) id NGPa017153 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Aug 1998 10:57:22 -0400 (EDT) Message-ID: {bc26a28c.35c72153-at-aol.com}
Dear all, I am placing this message for a third party, looking for a short course in electron microscopy preferably in regard to clinical pathology.
Unfortunately I missed prvious listings. Any information is wellcome.
Eckhart C. Dorneich Leo Electron Microscopy, Inc. ECDorneich-at-aol.com
Leica CLSM - confocal microscope (inverted). The system includes: a Leitz Fluovert inverted microscope, objective lenses, filters, krypton/argon laser, Motorola computer (running OS9) with Leica ScanWare software, and 3 monitors. Also for sale is a Focus Graphics image recorder.
If interested, please contact:
Scott Henderson, Ph.D. Director of Microscopy, Dept. Cell Biology & Anatomy, Mount Sinai School of Medicine, Box 1007, 1 Gustave L. Levy Pl., New York, NY 10029-6574
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } About staining with Reynolds lead citrate } } } } Put a clean 0.22 micrometer filter on the syringe before use. } } } Wet the filter with the solution before forcing a drop out } } } which is to be used for a grid. } } We have found that using a filter is not necessary. If the solution } has been made-up and stored properly it does not require filtration. } } } Hildy: I started dispensing lead citrate from a syringe recently, and find } } it works pretty well. However, would you clarify/elaborate the part 'Wet } } the filter...' ? } } As I reported earlier, we store the made-up Reynolds in approx. 3 ml } aliquots in Eppendorff tubes. We dispense by taking up the stain into } a new disposable Pasteur pipette from about 4 - 5mm above the bottom } of the tube and then discard the first drop. This minimizes the } possibility that any contamination from crystals, dirt, etc, which } may have accumulated at the bottom of the tube, on the surface of the } stain or in the pipette will end up in the staining drop. Once the } Eppendorff tube has been opened it is not re-used - it is discared } with any left-over stain. } } This uncomplicated method has produced almost totally } contamination-free stain for many years, even in the hands of the } most novice workers. } } Robin H Cross } Director : EM Unit, Rhodes University, Grahamstown, South Africa } eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 } http://www.ru.ac.za/affiliates/emu/em.htm } Sounds good to me! You have worked out a good method of making the stain which is of critical importance in the avoidance of precipitates. (By "wet the filter" I mean to drive a few drops of lead through the filter and discard these drops). If your method works, do not change it. I have heard of others who use Eppendorfs for storage successfully. (I am basically lazy (call it practical) and when I make up stain I make huge amounts like 500ml, and I distrubute that into 10 syringes, and I don't have to worry about it for 2 years). P.S. I hate making up stain. Bye, Hildy
NCEM is a national user facility housing state of the art electron microscopes and advanced image analysis and specimen preparation facilities, dedicated to the electron-optical microcharacterization of materials. The Center currently has an opening for a one year term Visiting Postdoctoral Scientist to: - Conduct research on Microstructural characterization of plastically deformed samples using the wide range of TEM facilities available at NCEM. Explore the correlation between local physical/chemical microstructure in plastically deformed regions with local changes in magnetic structure (measured independently by SQUID imaging techniques. - Develop Lorentz imaging methods on NCEM's CM200FEG microscope and appropriate computer modeling methods to interpret Lorentz images.
Qualifications: Ph.D.(within the last four years) in Materials Science/Physics or related area. Very strong hands-on experience in various TEM techniques and their application to microstructural characterization is required. Experience in Lorentz Imaging and TEM specimen preparation is desirable. Computing skills, particularly in developing/adapting code for micromagnetic modeling will also be helpful.
Send resume and cover letter to:
Dr. Kannan Krishnan, Lawrence Berkeley National Laboratory, One Cyclotron Road, M/S-72, Berkeley, CA 94720. Phone: 510/486-4614, Email: Krishnan-at-lbl.gov Fax: 510/486-5888
Good day everone. Does anyone know a current source for Janssen immunogold reagents? Thanks in advance. Robert Cox Shriners Hospital Burns Institute Galveston, Texas
Good day everone. Does anyone know a current source for Janssen immunogold reagents? Thanks in advance. Robert Cox Shriners Hospital Burns Institute Galveston, Texas
I am assuming the cells are fixed as a monolayer. I have sometimes had trouble with the osmolarity requirements of cell cultures. Check that. If they are a monolayer I would start with a more dilute fixative, such as 1 to 1.5 percent glut to help keep the osmolarity down, and you are only working with a few cells thick. We also add (10 percent by vol. ) saturated picric acid to stabilize membranes during aldehyde fixation. One investigator I worked with added 0.05M Potassium ferricyanide with the osmium to enhance neural tissue.(ref ?) If your fixative is refrigerated bring it to room temp so as not to shock the cells. Good luck with your cells.
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Date 8/4/98 Time: 6:08 PM Internal Memorandum
Does a commercial Thermal Wave Imaging Microscopy facility exist in the San Francisco Bay Area? If so please email me at Hwachob-at-exponent.com. Thanks you for your assistance.
by mail.exponent.com (8.8.5/8.8.5) with SMTP id SAA28489 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Aug 1998 18:14:07 -0700 (PDT) Message-ID: {n1309867756.63928-at-faamail.fail.com}
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Date 8/4/98 Time: 6:08 PM Internal Memorandum
Does a commercial Thermal Wave Imaging Microscopy facility exist in the San Francisco Bay Area? If so please email me at Hwachob-at-exponent.com. Thanks you for your assistance.
by mailhost.auckland.ac.nz (8.9.1/8.9.1/8.9.1-ua) with ESMTP id PAA27989; Wed, 5 Aug 1998 15:28:05 +1200 (NZST) Received: from MEDNOV1/SpoolDir by mednov1.auckland.ac.nz (Mercury 1.21); 5 Aug 98 15:27:25 +1200 Received: from SpoolDir by MEDNOV1 (Mercury 1.21); 5 Aug 98 15:27:24 +1200
} My colleague has had very poor TEM results with 2.5% glut in 0.1M } cacodylate with 0.1M sucrose, there are no membranes visible. Cell } membranes, mitochondrial membranes, organelle membranes, all } gone/invisible. Yes, post-fixation with osmium was used.
I suspect that the dehydration steps were too prolonged. For a thin layer of cells 3-5 minutes in each step should be plenty, and not too many steps ... e.g. 50% ethanol, 70%, 90%, 2 x absolute, propylene oxide, then resin.
} She has also had very poor SEM results with the same fix with the } addition of 4% paraformaldehyed, plasma membrane had large holes.
Sorry I can't offer advice on this, except to suggest using fixative at culturing temp (? 37 degC) and not ice-cold, and ensuring that the pressure changes in the CPD are not too sudden (both when first adding CO2 and at the end when returning to atmospheric pressure).
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
At 14:32 04/08/98 +0200, Krzysztof Jan Huebner wrote: } } } } Dear Friends, } } We have great pleasure to inform you about our conference "COLOR '99" } organised in Krakow under the auspices of Foundry Research Institute, } Krakow, Poland. } } Below we send to you the first announcement of this conference in the } file Word 7.0 "First.doc". } } } Best regards. } } } Members of Local Conference Committee: } } Janina Radzikowska and Krzysztof Jan Huebner } } } ================================================ } } } Krzysztof Jan Huebner }
I'm not interested by your conference but I received your attached file and it could be too much waste time for many people of the list server if everybody use this way each time there is a conference in the world. Please next time send only E.mail. Best regards Nicolas Stephant
We have great pleasure to inform you via e-mail about our conference "COLOR '99" organised in Krakow under the auspices of Foundry Research=20 Institute, Krakow, Poland.
Below we send to you the first announcement of this conference.
International Conference on Color Metallography Krak=F3w, Poland 19 - 21 May 1999
under the auspices of Foundry Research Institute Krak=F3w, Poland
Scope: This conference will be a meeting of scientists and industry workers interested in application of metallographic color techniques to=20 microstructural investigations of metals and nonmetallic materials with the use of the light microscope. The techniques of specimen preparation=20 will be presented and discussed as well. Exhibition of=20 instruments, equipment and photographic materials will make it possible=20 for the participants to get acquainted with the latest achievements in=20 the research and investigation technology. Conference'will be held in the= =20 MANGHHA Centre of Japanese Art and Technology in Krak=F3w with a beautiful= =20 view to Wawel castle.
Scientific Committee: G.F. Vander Voort (USA) - Chairman G. Petzow (Germany) P Skocowsky (Slovakia) K. Huebner (Poland) J. Radzikowska (Poland)
Main Topics of the Conference: 1. Preparation of specimens for color metallography 2. Light optical methods for color imaging: polarized light dark field differential interference contrast =B7 fluorescence 3. Color etching methods: tint etching anodizing interference layer method gas contrasting/reactive sputtering methods 4. Color image analysis procedure 5. Application of color metallography 6. Documentation of color images 7. New color techniques
Program: The scientific program will consist of keynote lectures, oral and poster=20 presentation and metallographic contest of color microphotographs.
Abstract submission: Abstracts should be in English and contain from 50 to 200 words on A4=20 sheet with wide margins. Along with the abstract, please indicate the=20 technical topic for which the abstract is being submitted; the title of the= =20 proposed presentation; and complete author information (name, address, phon= e, fax, E-mail) for all authors, with the presenting author listed first.=20 Alternatively, you can send MS Word 7 file (with True type fonts included)= =20 as an attachment to E-mail.
Before submitting your abstract or any other form of work listed in the=20 program, please be informed that all costs associated with participation=20 in the Conference will be at your expense, including travel, housing and=20 conference registration. Abstracts should be submitted to Conference Secretariat.
Deadline: 15 October 1998 for abstracts 30 January 1999 for papers 30 April 1999 for posters and contest microphotographs
Requirements regarding papers, posters and microphotographs as well as information referring to costs of participation and final program will be= =20 distributed later in the second announcement.
Conference language: English
Local Organizing Committee: Krzysztof Huebner Janina Radzikowska Halina Pawlowska Janina Danczak Krystyna Luszczkiewicz
Conference Secretariat: Krzysztof Huebner Foundry Research Institute Zakopianska 73 30-418 Krak=F3w POLAND Fax: (48-12) 266 08 70 Phone: (48-12) 266 50 22 ext.356 or 316 E-mail: hubner-at-iod Krak=F3w.pl.
Signature:
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D Krzysztof Jan Huebner=20
{hubner-at-IOd.krakow.pl} :-)=20
FOUNDRY RESEARCH INSTITUTE Research Materials Department Manager Structural and Physical Research Laboratory str. Zakopianska 73 Call (*48 12) 2665022 ext.356 30-418 KRAKOW - POLAND Fax (+48 12) 2660870
by jess.gla.ac.uk with esmtp (Exim 2.01 #1) for Microscopy-at-sparc5.microscopy.com id 0z430H-0002lD-00; Wed, 5 Aug 1998 13:49:17 +0100 Received: from l-tetley.chem.gla.ac.uk (l-tetley.chem.gla.ac.uk [130.209.221.186]) by lenzie.cent.gla.ac.uk (8.8.4/8.8.8) with SMTP id NAA27148 for {Microscopy-at-Sparc5.Microscopy.Com} ; Wed, 5 Aug 1998 13:49:15 +0100 (BST) Message-Id: {3.0.1.16.19980805134914.402fa156-at-udcf.gla.ac.uk} X-Sender: gbza40-at-udcf.gla.ac.uk X-Mailer: Windows Eudora Light Version 3.0.1 (16)
FOOD MICROSTRUCTURE - Towards the Year 2000 and Beyond
This conference is being held in collaboration with the Royal Microscopical Society
An understanding of the structure of foods and food products is essential to the efficient development of new processes or products. The future of the food industry is closely related to new developments in food microscopy.
This conference is an opportunity for those involved in food product development to hear the latest findings from key scientists in the field of food microscopy. The meeting will act as a forum to establish the needs of the food industry over the next decade, to discuss recent advances in food microstructural research and to evaluate new microscopical techniques.
This conference is aimed at food microscopists, food technologists and research scientists, particularly those involved in the development of new products or processes. Any company engaged in product development should be represented at the conference, to ensure that it is aware of the opportunities that advances in food microscopy can offer.
The aim of this conference is to:-
bring together food microscopists and food technologists to enable fundamental and applied research to be presented together;
highlight recent advances in food microstructural research;
provide a forum for discussion of microstructural studies relating to food;
identify the potential of new techniques.
Conference organisers: Mrs Kathy Groves and Dr Morag Saunders, Leatherhead Food RA; Dr Ashley Wilson, CCTR University of York.
There is still time for other papers to be submitted - please contact the conference organisers for further information.
KEYNOTE PAPER: Structure - the Only Thing that Matters? - Professor Peter Lillford, CBE, Unilever Research The challenge facing members of the food industry is to become architects of structure rather than processors of materials. This will need better application and novel approaches to microscopy.
Invited Paper: Cryogenic Environmental SEM of Ice Cream - an Introduction to the Principles of ESEM with Applications to Observations on Ice Cream - Brad Theil, Polymers and Colloids, Physics Department, University of= Cambridge
Food Microstructure using X-ray Projection Microscopy - John Judge and Peter Lillford
LUNCH
Chairman: David F. Lewis, SAC
The X-ray Microscope and its Applications to the Food Industry - Simon Burgess, Oxford Instruments
Probing Food Biopolymer Functionality with Atomic Force Microscopy (AFM) - Vic Morris, Institute of Food Research, Norwich (co-authors: A.P. Gunning, A.R. Kirby, A.R. Round, A. Mackie and P. Wilde)
The Environmental Scanning Electron Microscope - Speaker from Philips Electron Optics
Observations of Food Microstructure by Environmental Scanning Electron Microscopy - Debbie Stokes, Polymers and Colloids, Physics Department, University of Cambridge (co-author: A.M. Donald)
SESSION 2: APPLICATIONS OF MICROSCOPY TECHNIQUES IN FOOD RESEARCH
Chairman: Ashley Wilson, CCTR University of York
F Invited Paper: Applications of Variable Vacuum in Food Microscopy - Roger Angold, RHM Technology
F FTIR Microscopy in Troubleshooting for New Product Development - Hilary Holgate, RSSL
F Using Image Analysis and Confocal Microscopy Combined to Measure Deformation in Starch Matrices - Jeremy Addler, RHM Technology
F The Microscopy of Food. ESEM and Confocal Microscopy - Complimentary Techniques - Anthony Robinson, Polymers and Colloids, Physics Department, University of Cambridge
F Invited Paper: The Use of Electron Microscopy in the Investigation of BSE - Michael Stack, Veterinary Laboratories Agency
LUNCH and opportunity to view trade exhibition
SESSION 3: MICROSCOPY OF FOOD PRODUCTS, PROCESSES AND INGREDIENTS
Chairman: Roger Angold, RHM
Invited Paper: Microscopy - the Art of Food Technology - Professor Anne-Marie Hermansson, The Swedish Institute for Food and Biotechnology
Changes in Endosperm Structure during the Production of Popped Grain - Mary Parker, Institute of Food Research, Norwich
Confocal Laser Scanning Microscopy of Dairy Products and Ingredients - Methodology and Some Applications - Mark Auty, Dairy Products Research Centre, Moorepark, Ireland
Poster Exhibition and Opportunity to view Trade Exhibition
SESSION 3 (continued): MICROSCOPY OF FOOD PRODUCTS, PROCESSES AND INGREDIENTS
Chairman: Professor Anne-Marie Hermansson, SIK
Invited Paper: Brian Brooker, Institute of Food Research
The Use of SEM, Confocal Microscopy and Instrumental and Sensory Techniques in the Study of Biscuit Texture - Michael Edwards, Campden and Chorleywood Food RA
Crystallising Carbohydrates - Use of the Microscope in Understanding Crystallising Structures in Heat-resistant Chocolate and Powdered Sorbitol - Kathy Groves, Leatherhead Food RA
F Microstructural Changes of Food Products Following Inclusion of Non-starch Polysaccharides and Fibre Supplements: Implications to Dietary Nutrition - Karen Linton, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth (co:authors: C. Brennan and P. Moura de Magallraes)
LUNCH
Chairman: Brian Brooker, IFR, Reading; Mrs Kathy Groves, Leatherhead Food RA
Ultrastructural and Textural Changes in Heat-processed Fruits - Morag Saunders, Leatherhead Food RA
Cereal Structure: its Relationship to Raw Material Quality and End-product Utilisation - Charles Brennan, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth (co-authors: K. Linton, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth; D. Griggs, Pauls Malt Ltd; I. Cantrell, Pauls Malt Ltd; P.R. Shewry, IACR - Long Ashton)
Invited Paper: Perspectives on Food and Microscopy - David Lewis, SAC
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Are there other SEM, TEM, AFM,STM, EDS,WDS, Materials Analysis and Characterization Listserers? JD
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
I would recommend you to look at http://www.mwrn.com/ I found it to be very imformative in the area of microscopical resources
A ---------- =CE=F2: charles j day =CE=F2=EF=F0=E0=E2=EB=E5=ED=EE: 6 =E0=E2=E3=F3=F1=F2=E0 1998 =E3. 10:33 =CA=EE=EC=F3: Microscopy-at-sparc5.microscopy.com =D2=E5=EC=E0: Other EM and Materials Analysis Listserers?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Are there other SEM, TEM, AFM,STM, EDS,WDS, Materials Analysis and Characterization Listserers? JD=20
_____________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com Or call Juno at (800) 654-JUNO [654-5866]
If anyone has information or recommendations for embedding mud samples for TEM observation, I would appreciate it. To date, I have had some success with LR White resin in the microwave.
We are in need of a good atlas identifieng the ultrastructure of viruses. does anyone have any suggestions.
Christine Lee , Veterinary Pathology, University of Queensland Christine Lee, Veterinary Pathobiology, University of Queensland. C.lee-at-mailbox.uq.edu.au
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I'm looking for a method to stain tissue en bloc so that I can see it while I'm cryo-sectioning -- something with just a hint of color.
The tissue is frog saccule, which is small and so translucent that I can't see it as I approach it through the matrix, which is OCT. The tissue is destined for immunocytochemistry, so antigenicity needs to remain preserved. (It would be helpful to find a stain that may be used for other tissue types as well -- mouse retina and chick utricle, for example.)
I was told that methylene blue may be used, but I've been unable to find anything in my reference texts.
I would be grateful for any suggestions.
Thanks,
Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu
Center for the Study of the Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
by nss4.cc.Lehigh.EDU (8.8.8/8.8.5) with SMTP id PAA49232 for {microscopy-at-msa.microscopy.com} ; Thu, 6 Aug 1998 15:26:48 -0400 Message-Id: {3.0.1.32.19980806152641.006ca108-at-lehigh.edu} X-Sender: jae5-at-lehigh.edu X-Mailer: Windows Eudora Light Version 3.0.1 (32)
. I have a pair of glasses (polycarbonate lenses) which have become scratched. The opticians I have consulted say that there is nothing to be done except buy new lenses. The scratches are not deep. They are very fine but, since they are in the middle of the lenses, they render the glasses nearly useless.
Surely with all the expertise this community has in polishing materials of every kind, someone can tell me how to save $100.
Alwyn Eades . Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
This is a little off the topic, but microscopists who wear plastic lenses are bound to get them scratched. I have an answer for you, but it takes some elbow grease.
There are kits available for repairing CDs. Even though they may boldly say that they "fill" in scratches, the two that I have tried consisted of an abrasive solution and a soft pad. Make sure that you have removed all the dirt from your glasses and apply the opaque white liquid and rub with the pad. After a few minutes you should see the scratches "rub out."
I have even had moderate success restoring CDs!
Best regards from your token Optical Engineer mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
I know this has been covered in the distant past, but I never wrote it down. We have an open house coming up and I would like to have a micrograph from a CD for show and tell. Could someone tell me the sample prep steps to do this? I've never done it before.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Maybe you do require a printed atlas and somebody else will advise. However, there are many sites on the internet that feature virus and at least a couple with quite extensive collections. Go to our links page http://www.proscitech.com.au/links.htm and use the browsers control/F function to search for "virus" and "virol" Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Friday, 7 August 1998 2:44, Christine Lee [SMTP:C.Lee-at-mailbox.uq.edu.au] wrote: } ---------------------------------------------------------- } --------------
} We are in need of a good atlas identifieng the } ultrastructure of viruses. } does anyone have any suggestions. } } } Christine Lee , } Veterinary Pathology, } University of Queensland } Christine Lee, } Veterinary Pathobiology, } University of Queensland. } C.lee-at-mailbox.uq.edu.au }
I wonder if there is anyone working with EBIC who can provide me with = some advice. I have been asked to try and produce data from ZnO = varistors. In particular they want to relate the structure to current = flow across grain boundaries. I must point out that I am a complete novice to EBIC. I have obtained = a simple Specimen Current Monitor ( GW Electronics Type 31 ) which is = now connected to the microscope and producing EBIC images. The problem = is when I try to apply current biasing ( +/- 10V ), from an alkaline = battery, to a 5.5V device the only effect is banding/interference. = This may be due to a bad contact ( I will be making a new regulator ), = but there is no change in the EBIC image at the grain boundaries. I = have tried a range of KV's and probe currents without success. I would be very grateful if anyone can provide me with some advice.
Thanks,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.1712.3"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Hi, {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} I wonder if there is anyone working with EBIC who = can provide=20 me with some advice. I have been asked to try and produce = data from=20 ZnO varistors. In particular they want to relate the = structure to=20 current flow across grain boundaries. {/FONT} {/DIV} {DIV} {FONT size=3D2} I must point out that I am a complete novice to=20 EBIC. I have obtained a simple Specimen Current Monitor ( GW =
Electronics Type 31 ) which is now connected to the microscope and = producing=20 EBIC images. The problem is when I try to apply current = biasing (=20 +/- 10V ), from an alkaline battery, to a 5.5V device the only effect is =
banding/interference. This may be due to a bad contact ( I = will be=20 making a new regulator ), but there is no change in the EBIC image at = the grain=20 boundaries. I have tried a range of KV's and probe currents = without=20 success. {/FONT} {/DIV} {DIV} {FONT size=3D2} I would be very grateful if anyone can provide me = with some=20 advice. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Thanks, {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Colin {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Colin Reid, {BR} Electron Microscope=20 Unit, {BR} Trinity College Dublin, {BR} Dublin = 2, {BR} Ireland. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Tel: 353-1-6081820 {BR} Fax:=20 353-1-6770438 {BR} email: {A=20 href=3D"mailto:creid-at-tcd.ie"} creid-at-tcd.ie {/A} {/FONT} {/DIV} {/BODY} {/HTML}
Have a look in our links page http://www.proscitech.com.au/links.htm and find the category "Mail lists" using the find function. This section includes a couple of the listservers you are interested in. If others exist I would like to learn about these too. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Thursday, 6 August 1998 14:34, charles j day [SMTP:wa5ekh-at-juno.com] wrote:
} -------------. } } Are there other SEM, TEM, AFM,STM, EDS,WDS, Materials } Analysis } and Characterization Listserers? } JD } } __________________________________________________________ } ___________ } You don't need to buy Internet access to use free Internet } e-mail. } Get completely free e-mail from Juno at } http://www.juno.com } Or call Juno at (800) 654-JUNO [654-5866]
} I know this has been covered in the distant past, but I never wrote it } down. We have an open house coming up and I would like to have a } micrograph from a CD for show and tell. Could someone tell me the } sample prep steps to do this? I've never done it before.
We did something like this years ago. I can't put my hand directly on the records for the moment, but I think it went like this. The problem was that the dimples are not at the top of the CD, but buried inside with a metallic layer to reflect. As memory serves me, what we did was to cut out a small area of the CD, and then bed it shiny side down on some rapid cure Araldite. After this hardened, one could then flick off the piece of CD, leaving the shiny layer on the Araldite, and the dimples appeared as troughs in the surface that was exposed. This was then sputter coated with gold in the usual way, and examined under SEM with the direction of the grooves or dimple at the "magic angle" (put in flat, rotate so the the grooves run at 45^ to x and y, and then tilt z by 54^. I think one could also look at the remaining metallized area on the Aralidite, again gold coating.
There may be inaccuracies after such a long time of recall, but I think this will give you a general principle to work on.
Good hunting!
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Could you help me with ideas finding someone who would design a small lens system for a microscope/CCD relay lens? I'm in san Jose, California. Thanks! Julian Ortuondo
Hi Scott, I took a damaged writable cd and cut a wedge out, plastic and all. The foil peels off easily. I then mounted it on a stub with double sided carbon tab. The long line is called an "atip". This is the tract the lases follows. Then there are "lands" and "pits". These are the spaces(lands) between the laser burns(pits). Hope this helps.
Walck. Scott D. wrote: } . } } I know this has been covered in the distant past, but I never wrote it } down. We have an open house coming up and I would like to have a } micrograph from a CD for show and tell. Could someone tell me the } sample prep steps to do this? I've never done it before. } Regards, Gregory Rudomen Technical Specialist University Microscopy Imaging Center S.U.N.Y. Stony Brook 516-444-3126 Greg-at-umic.sunysb.edu
M. E. Haine, et. al. (Brit J. Appl. Phys. Vo,l 3, p. 40, '52; Vol 9, p. 482, '58; J. Brit. I.R.E. Vol. 17, p. 211, ' 57) made extensive studies of the effects of temperature, vacuum, and gun geometry on the life and brightness of tungsten filaments. This work is summarized in Ch. V! of Haine's book "The Electron Microscope", Interscience, 1961.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
Try 'painting' the suface of your specimen with either 1% methylene blue or 1% methyl green. Both are aqueous solutions.
-- Begin original message --
I'm looking for a method to stain tissue en bloc so that I can see it while I'm cryo-sectioning -- something with just a hint of color.
The tissue is frog saccule, which is small and so translucent that I can't see it as I approach it through the matrix, which is OCT. The tissue is destined for immunocytochemistry, so antigenicity needs to remain preserved. (It would be helpful to find a stain that may be used for other tissue types as well -- mouse retina and chick utricle, for example.)
I was told that methylene blue may be used, but I've been unable to find anything in my reference texts.
I would be grateful for any suggestions.
Thanks,
Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu
Center for the Study of the Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
-- End original message -- regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
I've lived with plastic lenses for decades. Any attempt at polishing wouldn't be worth the effort. The time you take, and the *extreme* care needed in polishing isn't worth effort. Remember, you not only have to eliminate the scratches, you have to retain your prescription. This can be difficult, especially if you have any prism or astigmatism corrections. Also the front and rear surface curvatures are not likely to be the same (you have concavo-convex lenses like most?), and the optical center is likely to be nearly as thin as the lenses can be safely made already. Particularly if they're safety lenses.
Only $100? (LeHigh doesn't pay for glasses?) If your lenses aren't all that thick, get glass lenses and lessen the scratching problem. Glass is cheaper, and you'll have to put off the getting the Miata for a shorter time. 8-)
Phil
} . } I have a pair of glasses (polycarbonate lenses) which have become } scratched. The opticians I have consulted say that there is nothing to be } done except buy new lenses. The scratches are not deep. They are very } fine but, since they are in the middle of the lenses, they render the } glasses nearly useless. } } Surely with all the expertise this community has in polishing materials of } every kind, someone can tell me how to save $100. } } Alwyn Eades } . } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvannia 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
I make a relay lens for microscopes and ccds. What are your requirements?
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
Dear Scott, I did this years ago, and as I recall, I cut a one centimeter square from the CD, put it in tri-chorethane overnight to dissolve the plastic, then fished out the very thin Al foil that was released and put it on a shiny graphite stub. It wrinkles horribly and the little grooves are hard to see until you reach 10,000X mag, but it is possible. This is for ressed CD's, the CD-R's you make yourself are completely different. You wrote: } } I know this has been covered in the distant past, but I never wrote it } down. We have an open house coming up and I would like to have a } micrograph from a CD for show and tell. Could someone tell me the } sample prep steps to do this? I've never done it before. } } -Scott Walck } } Scott D. Walck, Ph.D. } PPG Industries, Inc.
Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Alwyn, A fine metal polish: Brasso, Wenol, etc. should do the trick You wrote: } . } I have a pair of glasses (polycarbonate lenses) which have become } scratched. The opticians I have consulted say that there is nothing to be } done except buy new lenses. The scratches are not deep. They are very } fine but, since they are in the middle of the lenses, they render the } glasses nearly useless. } } Surely with all the expertise this community has in polishing materials of } every kind, someone can tell me how to save $100. } } Alwyn Eades } . } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Hello Again, Does anyone have experience with B-gal staining?=20 (5-Bromo-4-Chloro-3-indolyl-B-galactosidase), made in solution with 5mM potassium ferricyanide, 5mM potassium ferrocyanid= e and 2mM MgCl2 in PBS.
Specifically is it electron dense, or just something used at the LM? Thanks again, Linda M. Fox lfox1-at-wpo.it.luc.edu
Does anyone have a working recipe for Araldite 502?=20
Today I rec'd tissues, processed by another lab that followed a protocol = that stated "infiltrate in Araldite". They have the tissues in a single solution of Sigma A-3058, Araldite 502.
Any suggestions as how long to reinfiltrate in a complete recipe before = embedding? ( sm. int. aprox.1mm thick)
Also, we have 10 yr+ Araldite unopened in the lab. Does this go bad after = an extended shelf life??
Thanks as always... Linda Fox lfox1-at-wpo.it.luc.edu
A slightly different approach that I use is to mix a little stain (Toluidine Blue or whatever is handy) with the OCT. It's like negative staining if the tissue is light in color. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Hi-pressure bottled nitrogen for SEM venting is indeed the best straightforward solution. However, special precaution should be taken to avoid a pressure rise in the chamber, which may cause the implosion of the window of the EDS detector. I would like to draw your attention to a point that, if overlooked, may result in severe damage.
EDS detectors, with either Be or ultra thin polymer windows, will withstand a pressure difference of up to 1.2 bars, as declared by most maufacturers. In order to avoid the possibility of an accidental pressure rise in the chamber beyond this value we installed a sensitive pressure regulator, operable in the 0.05 - 2 bar range, on the nitrogen feed line. The salesman who sold us the regulator advised to set it to 1.2 bar, asuming that the pressure indicated is the absolute pressure. Unfortunately, this is not the case. The indicated pressure is the gauge (manometric) pressure, namely the pressure above the ambient pressure. Had we set it to 1.2 bar, the regulator would have allowed pressure build-up of 2.2 bar, which may ruin the EDS window. The correct value to set is 0.2 bar.
I do believe this seemingly trivial communication may be useful to some of you folks.
I am trying to do TEM on some marine sponges. They are looking at a symbiotic relationship with some algae . It was fixed in 2% glut dehydrated in ETOH and embedded in med - hard Embed 812. The problem is the tissue is tearing up the sections, due I suspect to the calcium spicules, to the point that I can't even get thick sections. I asked if we can decalcify the tissue first but they felt it might damage the tissue and the correlation of the two organisms.
My library access is medical so I turned up nothing on searches. 1. Has anyone had experience with sponges and TEM? 2. how about the effect on a diamond knife (sharpened for biological work)? Thanks for any help.
I forgot to mention that any polishing will also destroy any lens coatings, and all or almost all plastic lenses have such coatings-UV, anti-reflection, and anti-scratch.
Phil
} Alwyn, } } I've lived with plastic lenses for decades. Any attempt at polishing } wouldn't be worth the effort. The time you take, and the *extreme* care } needed in polishing isn't worth effort. Remember, you not only have to } eliminate the scratches, you have to retain your prescription. This can be } difficult, especially if you have any prism or astigmatism corrections. } Also the front and rear surface curvatures are not likely to be the same } (you have concavo-convex lenses like most?), and the optical center is } likely to be nearly as thin as the lenses can be safely made already. } Particularly if they're safety lenses. } } Only $100? (LeHigh doesn't pay for glasses?) If your lenses aren't all that } thick, get glass lenses and lessen the scratching problem. Glass is } cheaper, and you'll have to put off the getting the Miata for a shorter } time. 8-) } } Phil } } } . } } I have a pair of glasses (polycarbonate lenses) which have become } } scratched. The opticians I have consulted say that there is nothing to be } } done except buy new lenses. The scratches are not deep. They are very } } fine but, since they are in the middle of the lenses, they render the } } glasses nearly useless. } } } } Surely with all the expertise this community has in polishing materials of } } every kind, someone can tell me how to save $100. } } } } Alwyn Eades } } . } } Alwyn Eades } } Department of Materials Science and Engineering } } Lehigh University } } 5 East Packer Avenue } } Bethlehem } } Pennsylvannia 18015-3195 } } Phone 610 758 4231 } } Fax 610 758 4244 } } jae5-at-lehigh.edu } } }}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ } Philip Oshel } PO Box 620068 } Middleton, WI 53562 } (608) 833-2885 } oshel-at-terracom.net } or poshel-at-hotmail.com
Microscopy-at-MSA.Microscopy.Com I teach high school biology, anatomy and physiology and introductory bilingual life sciences. I have tried every source I can find to locate posters of microscopic iimages. Students are fascinated by them and as is well known, "a picture paints a thousand words". I have yet to be able to find a source of useful scientific images for my classroom. You only need so many nature posters. If anyone can direct me to a source I will be grateful. Robin Schaeffer Kodiak, Alaska 99615 honey1-at-ptialaska.net
Robin, I happen to have a series of 16 prints of high resolution electron microscope images by David Scharf. You may view them on my web site: www.microscopy- today.com Don Grimes, Microscopy Today
} On Fri, 7 Aug 1998, Linda Fox wrote: } } } Does anyone have a working recipe for Araldite 502? } } Yes but I can't get back to the lab for a few days. You } will need NMA, DDSA and DMP-30 for my recipe. } } } Also, we have 10 yr+ Araldite unopened in the lab. Does } } this go bad after an extended shelf life?? } } Mine didn't. } } Kal
But watch out for bad catalyst, particularly if you use DMP 30! You can assume that it's gone bad.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
One of my pet peeves...You can go out to any major bookstore and find a wealth of interesting material (books, magazines) about telescopes, astronomy, photography, however, try to find something about microscopy. At a number of stores you can find poor to pretty good telescopes. Celestron markets pretty decent telescopes in the $500 to $1000 range. Have you ever seen anything but a cheap plastic toy microscope sold in retail shops?
It seems to me that there could easily be a major consumer market for by periodicals and instruments that are good quality. Considering the relative simplicity of the microscope it should be possible to design a $100-500 microscope with metal base, glass optics, a substage condenser and iris diaphragm. A video camera attachment an i/o card to allow for computer capture and image manipulation would also be a relatively inexpensive affair.
Hi: A couple of years I was called to examine some CDs lookin for defects. I used 25% NaOH/DI water solution to disolve the aluminum layer, after one night the top plastic layer floated off and I careful decanted off the freed plastic layer. I then was able to gold coat the cleaned specimen and examine it succesufuly with our SEM. You can also remove most of the top coating with a solvent (I think I used 50/50 acetone toluene) which leaves the aluminum layer which can then be looked at (gold coating improves the resolution. Terry Ellis
Hi: A couple of years I was called to examine some CDs lookin for defects. I used 25% NaOH/DI water solution to disolve the aluminum layer, after one night the top plastic layer floated off and I careful decanted off the freed plastic layer. I then was able to gold coat the cleaned specimen and examine it succesufuly with our SEM. You can also remove most of the top coating with a solvent (I think I used 50/50 acetone toluene) which leaves the aluminum layer which can then be looked at (gold coating improves the resolution. I forgot! You do have to score a line around the edge of the CD to give the NaOH room to attack the metal layer. Terry Ellis
actually you can get an onld microscope like a B&L for theat price range and put the ccd camer on it. edmund makes a nice adaptor that will do the job.. Remember the old vcr recorders that used to sit on a strap and were sort of portable? They had seperate cameras that range from an easy conversion to a conversion via. hacksaw and some adaptor making. A pity you are not in Arizona, I have some old scope bodies that would work nicely for what you want to do..
I am going to also have some duplicate microscope books for trade and failing that I will sell some. Anyone that has ian interest drop me email. thanks Ed Sharpe Archivist SMECC
Hear hear! I couldn't agree with Jim more. I would love for my niece and nephew (and grandchildren when I have any) to be able to have a decent microscope of their own. They are at that age now where the mysteries of the world are becoming so inviting! It's a pity that there aren't any $100-$500 scopes available to kindle the interest of young minds in the science of microscopy.
} One of my pet peeves...You can go out to any major bookstore and find a wealth } of interesting material (books, magazines) about telescopes, astronomy, } photography, however, try to find something about microscopy. At a number of } stores you can find poor to pretty good telescopes. Celestron markets pretty } decent telescopes in the } $500 to $1000 range. Have you ever seen anything but a cheap plastic toy } microscope sold in retail shops? } } It seems to me that there could easily be a major consumer market for by } periodicals and instruments that are good quality. Considering the relative } simplicity of the microscope it should be possible to design a $100-500 } microscope with metal base, glass optics, a substage condenser and iris } diaphragm. A video camera attachment an i/o card to allow for computer } capture and image manipulation would also be a relatively inexpensive affair. } } Any entrepreneurs out there? } } Jim Harper
Jim -
There ARE good scopes and books out there! And the science store in the small town near me carries some of them; you must be unlucky. I can refer you to lots of mail-order sources; I have a page of phone numbers that I can Email to you (that page will appear on an expanded Project MICRO web page soon). Almost all of the scopes in that price range are Chinese, from a limited number of importers, but they're sold under lots of brand names.
Don't assume that a compound scope with condenser is the "best" for a beginner; a monocular dissecting scope is better for the youngest folks. You'll find detailed comments in MSA's new middle school manual, "Microscopic Explorations". Ordering info for it (and ~100 books, videos, & CD-ROMs) is in the MICRO bibliography - address below.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Ah, you are proving my point. There are obscure nooks and cranies that contain a wealth of information where microscopists can find them--but not in the main stream where the average consumer can find them.
I agree a stereo microscope is wonderful--probably my next major scope purchase--but can I find one at Nature's Wonders in the mall? (I can find at least 5 different telescopes there.)
I guess my point is that Nikon, Zeiss, etc. seem to be missing this market--or if there were decent microscopes offered in this price range would they no longer be able to sell a $10,000 stereoscope into the average business?
MICROFAB-at-aol.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Ah, you are proving my point. There are obscure nooks and cranies that } contain a wealth of information where microscopists can find them--but not in } the main stream where the average consumer can find them. } } I agree a stereo microscope is wonderful--probably my next major scope } purchase--but can I find one at Nature's Wonders in the mall? (I can find at } least 5 different telescopes there.) } } I guess my point is that Nikon, Zeiss, etc. seem to be missing this market--or } if there were decent microscopes offered in this price range would they no } longer be able to sell a $10,000 stereoscope into the average business? } } Jim
Jim,
We are trying to produce consumer level microscopes, but interest (and sales are slow) and this makes no economical sense for us. Check out the new Naturescope; it is a stereo for under $400......
} We are in need of a good atlas identifieng the ultrastructure of viruses. } does anyone have any suggestions. } } } Christine Lee , } Veterinary Pathology, } University of Queensland } Christine Lee, } Veterinary Pathobiology, } University of Queensland. } C.lee-at-mailbox.uq.edu.au } } } ---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: GPO Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 (0)2 6249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm
As many subscribers probably are aware, the Microscopy Society of Southern Africa (MSSA) is bidding to host the 15th International Congress on Electron Microscopy (ICEM) in Durban, South Africa, from 1 - 6 September 2002.
Presentations by societies bidding to host the 15th ICEM, and voting on the venue, takes place during the 14th ICEM in Cancun, Mexico, later this month.
Anyone wishing to have information on the Southern African proposal is invited to visit our web site (http://www.ru.ac.za/affiliates/emu/icem2002.htm) and associated links, or may contact me directly.
Those attending the 14th ICEM in Cancun are welcome to visit the Microscopy Society of Southern Africa's booth where information will be available about the bid for the 15th ICEM as well as other information about the Society and microscopy in general in Southern Africa.
Looking forward to meeting as many as possible of you in Cancun.
Best regards,
Robin Cross Vice-President : MSSA
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
I am looking for an antibody against mouse! alpha-fetoprotein. We would like to perform immunohistochemistry on mouse tissue. Has someone made positive experiences with any antibody against this protein?
I have received over the e-mail what purports to be a TIFF file of an AFM image. However, it has been "7 bitted" for transmission using something called Bin Hex 4.0, (on a Mac, I think), and needs to be decoded using the same. Does anybody know about this beastie, and if it can work on a PC?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I agree that the relative cost of a microscope for kids is high. For example, I can buy a pair of binoculars for $25, which contains more than twice the optics (all achromatic glass elements) and roughly the same mechanical components as a beginners compound microscope. The difference is that there is a big market for binoculars, and not for microscopes. One idea might be to develop a kit for converting a pair of cheap binoculars into a microscope (see patent 3,804,486 by Gerrit A. Van Extl and Alfred A. Akin, Jr.; assigned to Bausch & Lomb).
However, you can still buy a good compound microscope (achromatic glass elements, DIN objectives and eyepiece) for under $150 from a well known mail order scientific equipment company. I have used this scope with kids and it is great.
Several years ago I had a month-long microscope show at my local science center with the goal of getting people to buy low-cost microscopes for their kids. I felt that the cost of the microscope was a definite hurdle, but more significant was the lack of good books describing microscope activities for kids. Most (but not all) of the books readily available are plain stupid and not helpful. Interestingly, my local library has an excellent technical section with a number of turn-of-the-century books on microscopy (Marvels of pond-life; or, A year's microscopic recreations among the polyps, infusoria, rotifers, water-bears and polyzoa by Henry J. Slack, 1897 and Hunting under the microscope by Sir Arthur E. Shipley, 1928). Microscopes would be more popular if books like these were available today.
Everett Ramer Federal Energy Technology Center Pittsburgh, PA
While my EM Unit caters strictly to research, I am often asked whether I would agree that the usefulness of the EM in the clinical setting is in a major decline. It is pointed out that diagnostic EM, especially in the area of tumors, is rapidly being replaced by immunocytochemistry at the LM level. In addition, tumor cells do not as a class exhibit ultrastructural changes that can tell us much in the way of significant information anyway. I was wondering if you might have an opinion on this observation or a reference(s) where I could check this out.
Hi..having a problem with powder examination; have never before looked at powders (TiO2 in this case). Used conducting carbon adhesive pads to fix sample to stub but not much stayed on and although it coated ok most of sample came off whilst pumping out in the sample exchange chamber of our Hitachi S-800...How can i get round this..is there some sort fo fixative spray? Cheers Alan. Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
My Leica paraffin tissue embedding station (essentially a hot wax dispenser) has a broken heating plate and Leica wants $3600 to repair it. I can get a new Leica for $6400 but would like to consider alternatives. Does anyone know of a different brand? I don't need a tissue processer (to run the specimen thru fix/rinse/ethanol/xylene and infilitrate in wax). I just need something for the final placement of tissue into molds. TIA. Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Linda We have been using this resin in the research lab and up in the path EM lab for over 15 years. Their formulation was based on Luft's work I believe. It is one of the most viscus resins so we take a long time infiltrating. Our last dehydration / dealcoholization step is propylene oxide "PO" (yeah I know it's toxic) 3x 10 min. we then replace with a 50/50 mix of complete "comp" resin and PO The initial resin mix is; (27 ml 502, 23 ml DDSA. if you warm the bottles to 40-50 degrees C it stirs easier). From this mix an aliquot is poured out and a 0.2%volume of DMP-30 is mixed in. We call this "comp". (Some people substitute BDMA for the DMP30 but I didn't see an infiltration or cutting advantage and you sue twice the amount as DMP30.) This 50/50 mix is left capped over night. The next day we replace the 50/50 with 100% fresh comp for two hours, then embed into gelatin capsules or flat molds. The 100% infiltration and embedding resin should be made up that day not the previous days resin (it can be stored in the fig. and used for 50/50 the next time, if that is not too long, say a couple weeks.)
In regards to OLD DMP-30.......they use it up pretty fast but buy it volume so the last bottle is probably a year old. It is an anhydrous chemical and the suppliers suggest a 3 month shelf life but obviously it can be extended. I guess like some of the other topics we discuss ie stain and fixative life its how much work vs precision, and whether you can redo a grid or experiment. We're good but usually not picky, if in doubt order fresh stuff its cheap. Lately I have switched over to the epon 812 group of resins, as they are much less viscus and cut just as well. Good luck with your work.
The Call for Papers for the International Conference on Metallurgical Coatings and Thin Films -1999 in San Diego is out. Details of the meeting can be found at the following web site.
http://home.vacuum.org/icmctf/icmctf.html
Of specific interest to physical science researchers on the Microscopy Listserver is Symposium F. There is an emphasis this year on scanning probe microscopy techniques. Session F4/B3 concentrates on Microstructural, Microanalytical and Imaging Characterization of thin films. I've included the Symposium F and the F4/B3 session synopses below. Submission of abstract can be done over the Internet.
Symposium F Coating and Thin Film Characterization Symposium Chairs: John T. Grant, Research Institute, University of Dayton, Dayton, OH 45469-0168 Phone: (937)255-6603; Fax: (937)258-8075; e-mail: grantjt-at-ml.wpafb.af.mil Hans J. Steffen, Mannheim University of Applied Sciences, Mannheim, Germany Phone: (49)621-292-6543; Fax: (49)621-292-6420; e-mail: steffen-at-fh-mannheim.de
http://home.vacuum.org/icmctf/symposium f.html
OBJECTIVES: This symposium focuses on applications and recent advances in coating and thin film characterization. Moreover, it deals with progresses in the fundamental understanding of film growth processes and the elementary structure - properties relations by analytical methods, especially but not exclusively in the fields of carbide, oxide, nitride, and DLC coatings. It is the intention of the F symposium to create an analysis group which works on hard coatings and deposition techniques for the discussion and exchange of ideas and procedures for the elucidation of growth processes and mechanisms. These topics are of particular importance as emerging deposition processes and innovative processing techniques are employed to produce thin films and coatings with unique mechanical, chemical, physical, and microstructural characteristics. Of special interest are analytical and characterization techniques and methods, including numerical evaluation procedures like factor analysis etc. to adequately describe these coatings and thin films during and after the deposition. This symposium also addresses the unique analytical challenges in the investigation of functionally gradient, multilayer, nanocrystalline, heterogeneous and composite coatings. Nondestructive and in situ characterization of all kind of coatings are also of special interest. Hence, a particular focus will be on X-ray diffraction analysis.
In 1999, Symposium F will highlight applications of all scanning probe microscopy techniques (AFM, STM, etc.) and imaging methods. These techniques have wide applicability for characterizing state of the art coating materials and thin-film architectures. The session includes topics covering theory, experiment, and sample preparation. Submissions are welcome on characterization of: mechanical, chemical, and structural properties of thin films and coatings such as friction, wear, adherence, topography, roughness, hardness, phases, and chemical forces.
Invited Speakers: Michael Serry, Digital Instruments,Recent Technologies and Advances in Thin Film Characterization Using Atomic Force Microscopy and K. Satori, Sony Corporation, Japan,Surface Roughness Development During Sputter Depth Profiling of Semiconductor and Metal Thin Films determined by AFM
F4/B3. Microstructural, Microanalytical and Imaging Characterization. Session Chairs: Siegfried Hofmann, National Research Institute for Metals, Japan and Scott D. Walck, PPG Industries. This joint session solicits papers covering advanced characterization of the micro- and nano-structure of coatings and thin films. This session will focus on recent developments in new and established techniques for microstructural characterization and imaging, with emphasis on surfaces and interfaces. Methods include but are not limited to high resolution and analytical TEM, STM/AFM, EPMA, XRD, SEM, grazing X-ray reflectivity (GXR) and other spatially resolved imaging and elemental mapping techniques. Invited Speakers: Isao Kojima, National Institute of Materials and Chemical Research,High Resolution Thickness and Interface Roughness Characterization in Multilayer Thin Films by Grazing Incidence X-ray Reflectivity, Larry F. Allard, Oak Ridge National Laboratory,Materials Characterization Via the Internet, and Kannan Krishnan, Lawrence Berkeley National Laboratory,Magnetism and Microstructures in Thin Films, Multilayers and Nanostructures.
Hello.......we are starting to do Fluorescent In Situ Hyb's of different bacterial species. My basic question is, can I re-use the slides? I use Cell-Line teflon coated slides with 6 wells, that have been cleaned with 10% KOH and then subbed with 0.1% gelatin and 0.01% chromium potassium sulfate.
Thanks, Chris Odt ---------------------------------------------- Chris Odt U.S. Dairy Forage Research Center, USDA-ARS 1925 Linden Drive West, Madison WI 53706 FAX 608-264-5147, Phone 608-264-5320
My Life According to Molecular Biology : DNA} RNA} Protein} Traits} One Spirited Red-headed Daughter
email: clodt-at-facstaff.wisc.edu "Live as though your hair is on fire....even if its thinning" Ann Onymous ----------------------------------------------
You can get some very nice posters/calendars from a number of vendors that have beautiful micrographs in them. Consider contacting Gatan, Buehler, Nikon, Olympus, Leica, Polaroid, and the different EM manufacturers for some. (This is not a complete list, but some that I know have had these in the past.) Ask them if they might have a collection of old posters and calendars as well as the current crop. I think that you will be very impressed with what some of the vendors have.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Microscopy-at-MSA.Microscopy.Com I teach high school biology, anatomy and physiology and introductory bilingual life sciences. I have tried every source I can find to locate posters of microscopic iimages. Students are fascinated by them and as is well known, "a picture paints a thousand words". I have yet to be able to find a source of useful scientific images for my classroom. You only need so many nature posters. If anyone can direct me to a source I will be grateful. Robin Schaeffer Kodiak, Alaska 99615 honey1-at-ptialaska.net
Alan, You didn't say what you are examining the powders for, but here are a = couple methods we use for powders: 1. We use a clear double-stick tape (about =BC" wide). Cut a small = piece of tape, and stick to s sample stub. Peel off the top masking layer of = the tape, and press the stub (tape side first) into a sample of the powder. Lightly blow off the stub w/ filtered dry N2 to remove loosely adhered particles. Gold coat (15-20 nm) and examine. We haven't had good luck = w/ the adhesion of the carbon pads.
2. If you are doing feature analysis on the particles and don't want agglomerations of particles, we mix 0.5 gm of sodium pyrophosphate in 1 liter of DI water. This mixture has a shelf life of 24 hours. Put approximately 0.02 gm of powder (a small bit on the end of a tooth = pick) into 40mL of this solution. Mix solution very well and withdraw a = small amount using a syringe. Make sure you mix well to keep larger = particles in suspension. Deposit a drop of the solution onto a highly polished = stainless steel coupon. Evaporate the water in an oven at 100F and examine. The sodium pyrophosphate serves as a dispersion agent to reduce particle agglomeration. You probably won't have to gold coat the sample with = this method.
John Giles Principal Materials Engineer Honeywell Space Systems Clearwater, FL
} Hi..having a problem with powder examination; have never before looked = at powders } (TiO2 in this case). Used conducting carbon adhesive pads to = fix sample to stub but not } much stayed on and although it coated ok most = of sample came off whilst pumping out in } the sample exchange chamber of = our Hitachi S-800...How can i get round this..is there } some sort fo = fixative spray? } Cheers } Alan. } Dr Alan Templeton } Centre for Physical Electronics and Materials } SEEIE
Beta-galactosidase cytochemistry (with X-gal) is usually done at the LM level. However, here are some references in which the activity is viewed by electron microscopy:
Engelhardt JF, Allen ED, Wilson JM, 1991. Reconstitution of tracheal grafts with a genetically modified epithelium. Proc Natl Acad Sci USA 88:11192-11196.
Franklin RJM, Barnett SC, 1991. The electron microscopic appearance of the beta-galactosidase reaction product. Acta Neuropathol 81:686-687.
Liu HS, Cardell, EL, Stambrook PJ, Cardell, RR, 1991. Bacterial beta-galactosidase expression in cultured mammalian cells: light and electron microscope analysis of epon sections. Anat Rec 229(4):54A (abstract).
Loewy AD, Bridgman PC, Mettenleiter TC, 1991. Beta-galactosidase expressing recombinant pseudorabies virus for light and electron microscopic study of transneuronally labeled CNS neurons. Brain Res 555:346-352.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
} Does anyone have experience with B-gal staining? (5-Bromo-4-Chloro-3- } indolyl-B-galactosidase), made in solution with 5mM potassium } ferricyanide, 5mM potassium ferrocyanide and 2mM MgCl2 in PBS. } } Specifically is it electron dense, or just something used at the LM? } Thanks again, } } Linda M. Fox } lfox1-at-wpo.it.luc.edu
Scott- previously I produced SEM samples of CD's to evaluate the etching or burning from the laser involved in the writing process. simple steps (as long as you understand the CD will be destroyed) immerse the CD in liquid nitrogen (other cryo-liquids will probably work also) for approx. 30-60 sec. You will hear it start to crack. Remove the CD with tongs, slap it down onto a hard surface (lab bench, desk) or strike it while lying on the hard surface with a hammer. The plastic/metal interface will sheer, leaving a conductive metal to examine under the SEM. I usually used carbon sticky tabs to mount, no sputtering was necessary. Good Luck -Mike
You first need to determine the composition of the spicules. Are they only calcium? Many groups of sponges have both calcareous and silaceous spicules. If they are indeed only calcareous, I'd try one of the block-face decalification procedures. These can be found in the histology and especially bone and tooth histology literature, or email me and I can give you the address of a person or two who does this.
The diamond knife might work, but you likely will need to get the knife resharpened. Ask them if they need TEM sections...perhaps SEM would answer the questions they're asking as well or better.
Phil
} } I am trying to do TEM on some marine sponges. They are looking at a } symbiotic relationship with some algae . It was fixed in 2% glut } dehydrated in ETOH and embedded in med - hard Embed 812. The } problem is the tissue is tearing up the sections, due I suspect to the } calcium spicules, to the point that I can't even get thick sections. I asked } if we can decalcify the tissue first but they felt it might damage the } tissue and the correlation of the two organisms. } } My library access is medical so I turned up nothing on searches. } 1. Has anyone had experience with sponges and TEM? 2. how about } the effect on a diamond knife (sharpened for biological work)? } Thanks for any help.
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{{{{{ Philip Oshel PO Box 620068 Middleton, WI 53562 (608) 833-2885 oshel-at-terracom.net or poshel-at-hotmail.com
Linda- we currently use this recipe: Araldite 502 15ml Eponate 12 (or equivalent) 25ml DDSA 55ml DMP-30 (catalyst)(1.5%) 1.25ml
there are several mixtures (of varying hardness) in print, check any Hayat or Glauert book on basic principals in EM. as for reagents that are 10 years old (this stuff is cheap, your time is not) buy new resins, plastics, DMP-30, etc. -Mike
On Fri, 7 Aug 1998, Linda Fox wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Microscopy Friends, } } Does anyone have a working recipe for Araldite 502? } } Today I rec'd tissues, processed by another lab that followed a protocol that stated "infiltrate in Araldite". } They have the tissues in a single solution of Sigma A-3058, Araldite 502. } } Any suggestions as how long to reinfiltrate in a complete recipe before embedding? ( sm. int. aprox.1mm thick) } } Also, we have 10 yr+ Araldite unopened in the lab. Does this go bad after an extended shelf life?? } } Thanks as always... } Linda Fox } lfox1-at-wpo.it.luc.edu }
A faculty member here has asked me to look for images of 'aquatic pathogens' he may use on his teaching web site. He has mentioned such things as E. coli and V. cholerae but would like as great a range as possible. He wants pictures that he can use without problems of copyright releases etc.
He is also interested in other web sites or 'distance learning' resources that might 'provide the students with additional exposure to pathogenic microorganisms'. Personally, I try to avoid pathogenic microorganisms, but students may need the exposure.
Reply to me if you have something that could help him and I will pass along your offers.
Thanks
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu **Area code changing to 831 as of 7/11/98**
Alan Templeton wrote: ================================================ Hi..having a problem with powder examination; have never before looked at powders (TiO2 in this case). Used conducting carbon adhesive pads to fix sample to stub but not much stayed on and although it coated ok most of sample came off whilst pumping out in the sample exchange chamber of our Hitachi S-800...How can i get round this..is there some sort fo fixative spray? ================================================= Having looked at a lot of powders of this type ourselves, the first question is this: How old are the "carbon adhesive pads"? People often times don't realize it, but they do have a shelf life and with the passing of time, they do lose some of their original "tac". The temperature of their storage can have a lot to do with the rate of deterioration of the "tac", stored in a refrigerator, they will last much longer than if at room temperature.
Good fresh double sided conductive carbon sheets and tapes, such as you would find on the websites of SPI and some of our competitors, should contain sufficient "tac" to hold the particles. When applied dry, we like to apply a few "blasts" from a "duster" which serves to blow off the excess and also to embed a bit more firmly into the adhesive, the particles you want to study.
Another way is to mount the powders on our Tacky Dot™ Slides. See our website below for details. The smallest dot size presently available is 15 µm and if this is more than 4X larger than the size of the powder you have, the worst thing that will happen is that you will have some doublets and triplets on the dots. However, even if there should be more than one particle per dot, it is still a better way, often times, to characterize this kind of a powder, especially when using stage automation for large numbers of samples.
Disclaimer: SPI Supplies manufactures Tacky Dot™ Slides under license from the DuPont Company so we would have a vested interest in seeing more of them in use! And we also offer double sided adhesive conductive carbon sheets and tape as indicated above.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I am looking for a horizontal detector to fit a JEOL 100CX-II with an ASID or I need to get a Hack-Z package so I can install the high angle detector I currently have. Any help I get with respect to this matter will be greatly appreciated. Thanks, Mike Pidgeon
I have a JEOL 100CX-II with an ASID and need to generate some income with it. Preferably something that does not require accreditation. Does anyone have a suggestion on what I might do, and do you possibly have some contacts. I would appreciate any input on this subject. Thanks, Mike Pidgeon
Unlike a previous poster, I have had good luck using carbon loaded double face (actually mastic only) tape. With a finely divided insulator like TiO2, a thin well adhered layer is important so as to not break conduction paths after the conductive film has been applied. I prepare the mount by (as previously mentioned) gently thrusting the tape/mount into the powder or by "dusting" it onto the tape surface. Usually I don't blow off the excess, but instead sharply tap the mount a number of times while it is held up side down in my fume hood (many of my samples are rather "nasty").
I then will sputter with Au if well resolved morphology is the goal. If BSE and x-ray analysis is required, I (yarn) evaporate carbon instead. To meet both goals it is sometimes necessary to prepare two samples, one sputtered , the other C-evaporated.
...The Etec vacuum controller normally slams open a large gate valve causing a rush of air (N2 really) out of the chamber. Before modifying the pump-down, I lost a few 10mm cubes of fluffly ceramic insulation down the tubes:( I have installed a manual vacuum valve on my Etec so that I can gently rough
pump the chamber. This reduces the likelyhood that the coating will be corrupted.
Woody White McDermott Technology, Inc. mtiresearch.com
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Hi..having a problem with powder examination; have never before looked at powders (TiO2 in this case). Used conducting carbon adhesive pads to fix sample to stub but not much stayed on and although it coated ok most of sample came off whilst pumping out in the sample exchange chamber of our Hitachi S-800...How can i get round this..is there some sort fo fixative spray? Cheers Alan. Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
by sciences.sdsu.edu (8.8.8/8.8.8/SCEC-8.8.8-AS.AR.S5) with ESMTP id OAA01654 for {microscopy-at-msa.microscopy.com} ; Mon, 10 Aug 1998 14:27:50 -0700 (PDT) Message-Id: {v03020902b1f517bb3299-at-[130.191.234.197]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
howdy all
anyone who knows where I can get a 35 mm camera for a Philips 410 please let me know offline
thanks
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Gary Take a look at the books by Dvorak and Monahan-Earley "Diagnostic Ultrastructural Pathology I, II, and III" (CRC Press 1992 ISBN 0-8493-4404-2) and then consider the usefulness of EM in diagnostic medicine.
Bob
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718
For those interested, you can get a short term internet access account via Mr. Gaston C. Blanc at {gastonb-at-acnet.net} in Cancun. The charge is US $50 for the short term account which includes 150 hours of use. I have been told that payment is to be by cash only and that the service has to be arranged for in advance (but paid for in Cancun when you get your account set up).
There don't seem to be any local Compuserve numbers in that part of Mexico and calling the USA from there is expensive.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Greetings to all from the Inter/Micro 98 Meeting in Chicago. As some of you will remember, last year I made daily posts to this list summarizing some of the interesting papers being presented at Inter/Micro. This year I will again provide daily summaries but they will appear at the newly created MSA "Email-to-WWW Announcement Page" located at http://www.msa.microscopy.com/WWWviaEmail/EmailtoWWW.html . Please go there on a daily basis to review notes on many of the fascinating papers being presented at this, the 50th Anniversary Inter/Micro Meeting. Congratulations to Dr. Walter McCrone and all of the staff at McCrone Research Institute for one half century of fantastic meetings. May we live to see as many more!
I was wondering whether anybody has any experience with pointed W filaments in a XL-30. Ours currently has a normal W, although it is design for LaB6 operation. The problem I have with LaB6 is poor stability due to contamination build up on the Wehnelt (vac~2.5e-7 mBar) and hence short operation time between cleaning (not filament life time). The W is nice and stable and has been operating for several months without a clean, however I need a bit more brightness.
Are there any advantages to using pointed filaments over normal? What are the disadvantages? Are they worth the extra trouble?
Keith.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
I am a novice in this field, but have already made progress in this field specifically on polished sections of clinker specimens using etching and the like. We have a MEIJI microscope from which I can gather does not have excellent optics. Does anyone know anything about this brand of microscope and is it worth setting up a digital camera for it to do image analysis. Also we want to get a point counting stage. Any recommendations would be very welcome. Thank you.
Sincerely, Quirina
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
Pointed filaments have less than half the life, on average, when compared with standard filaments. They are more expensive and cause more downtime. They were one of the few means to achieve that extra little brightness 20 years ago. LaB6 give more brightness and reduce downtime a great deal. In the end, they are economical. Your Wehnelt contamination problem may be related to the temperature of your cooling water (Hong Kong in summer) or a number of other reasons. I would try to improve the vacuum. I also note that Kimball Lab6 cathodes arguably have the most robust design and can stand more overheating occasionally to remove contamination. I must declare that ProSciTech provides Kimball Physics Lab6 cathodes. Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Tuesday, 11 August 1998 17:51, Keith Moulding [SMTP:mcmouldk-at-uxmail.ust.hk] wrote: } } Greetings, } } I was wondering whether anybody has any experience with } pointed W } filaments in a XL-30. Ours currently has a normal W, } although it is design } for LaB6 operation. The problem I have with LaB6 is poor } stability due to } contamination build up on the Wehnelt (vac~2.5e-7 mBar) } and hence short } operation time between cleaning (not filament life time). } The W is nice and } stable and has been operating for several months without a } clean, however I } need a bit more brightness. } } Are there any advantages to using pointed filaments over } normal? } What are the disadvantages? } Are they worth the extra trouble? } } Keith. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Dr. K. Moulding. } } Materials Characterisation and Preparation Facility } Hong Kong University of Science and Technology, } Clear Water Bay, } Kowloon, } Hong Kong. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ }
Energy Beam Sciences developed a range of pointed tungsten filaments more than 25 years ago, and we are certainly the primary source of these filaments worldwide, so my bias should be obvious. However, I am a professional manager, not a salesperson, so I will simply tell you what I know.
All pointed filaments are not created equal. True pointed filaments consist of single crystal tungsten points welded on top of a filament loop. While the emission is from the point, much higher brightness can be acheived. However, there are two caveats:
1. A clean vacuum is required. The point will burn out almost instantly under poor vacuum conditions.
2. Under the best of circumstances, pointed filament life is relatively short, because of the very small emission point. Once there is suffiecient material loss from the point, the filament behaves as an ordinary "loop."
Intermediate between standard filament loops and true pointed filaments is a class of filaments etched back at the tip to create a "scalpel" shape. These filaments will not be as bright as the true pointed filaments, but they are more stable, and filament life will be close to normal. We always advise customers new to pointed filament to try these first.
Please contact me back-channel if you would like additional information.
Best regards, steven E. slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Having just gotten back into the SEM community following a 12-year hiatus, and while trying to get back up to speed by browsing through the archives of this Microscopy list server, I read with extreme interest the articles posted recently on the subject of the "ADEM1." In that thread, Craig Theberge gave a partial answer from his own personal experience to the question(s) that were raised, but I feel compelled to fill in more of the historical details, completing the picture from my own (albeit unique) perspective...
ADEM was the internal code name for a major new secret project which Fred Schamber headed up starting in 1985. (ADEM stood for "Advanced Digital Electron Microscope", I believe, but was merely coined as a code name and was not initially intended to be the actual name of the product.) The development team was organized as a separate company (later to be known as Tracor EBI) with it's own multi-million-dollar budget and staff (consisting of only a handful at the onset, handpicked from the Tracor-Northern R&D staff.) Temporary facilities were rented off-site in the Middleton Industrial Park (later dubbed the "Skunkworks") to house the entire top-secret operation. We ended up staying there for the next several years and even expanded the facility (renting adjacent bays) to accommodate additional staff (adding on to engineering, but also office personnel, stock, purchasing, drafting, etc.) Most people at the main Tracor-Northern plant didn't know what was going on just across town, and many didn't even know that the facility even existed. It was being kept a secret for the reasons that Craig mentioned - as a key microanalysis player at that time, we couldn't afford to lose our alliances with existing SEM manufacturers until we had perfected our own column. Fred was really going out on a limb with this, putting the company's future at risk financially, but felt he had the technical expertise to pull it off, and corporate was willing to trust him because of his many previous successes. After all, it would not have been an exaggeration to say that Fred was largely responsible for building the success that TN enjoyed by 1985, with a series of successful products dating all the way back to the NS-880.
Of course, in view of the unprecedented financial investment involved, Fred was under considerable pressure to deliver a return on investment in timely fashion and thereby prove that corporate's confidence in him had not been misplaced. The core R&D personnel involved at the inception were also quite dedicated to the success of the project, and so you can bet that for the next couple years, there were plenty of late hours and weekends devoted by all. Most of these have gone their separate ways now, but they included Mike Krummey (electronics engineering), Jorgen Rasmussen (mechanical engineering) and myself. We also had two primary consultants on the project: Dave Pearson (an experienced Etec serviceman, who brought in an old junker Etec scope which he kept running for us to use as a test bed for proof-of-concept on several hardware design issues), and Rich Lee! Yes, that's the same Rich (R.J.) Lee whom Fred later joined to produce the Personal SEM. A few other individuals were brought in from the outside as the need for certain skills became apparent - Ken Spielman, for example (who is still at Noran, by the way), was a skilled machinist hired in from the UW to hand-machine all the various sundry precision metal parts needed in the construction the first prototype. And of course there were technicians, draftsmen, purchasing and office support personnel, etc., added as time went on (and even marketing, sales and service people still later.) But it was Fred who spearheaded and masterminded the whole operation. It was his dream initially, his brilliance as a physicist which was crucial in solving several key problems along the way, his resourcefulness which kept the project on budget, and it was his drive and determination which kept the project moving ahead.
Fred had his hand in nearly every aspect of the development, but soon was spending the majority of his time focusing (pun intended) on the design of the lenses, which involved sophisticated electromagnetic field computations as well as careful selection of appropriate alloy materials for the pole pieces, etc. But despite the complexities involved in that, and with all due respect to Fred, those were among the more conventional aspects of the ADEM. What made ADEM truly unique was its huge chamber, sophisticated and fully-articulated sample stage, along with complete integration of all subsystems and digital control of each utilizing distributed microprocessor control. It was my pleasure to develop software and firmware for several parts of the latter, including the entire six-axes stage controller (consisting of microprocessors to drive each of the three translational and three rotational axes, and a "master" processor to coordinate the motions of all six axes.) In fact, microprocessors drove every subsystem of the ADEM, including not only the stage, but also the scan generator, vacuum system, EDS front end, etc., even the water valve! Dear reader, please indulge me as I proudly mention that I was responsible for the embedded software on nearly all of these subsystems, and personally wrote the code for most of them. But the whole instrument was tied together by a central processor which also performed the primary imaging functions; and the software for that part of the system was developed by a team of several other talented programmers. Of course, we were working with several bright electronics and mechanical engineers who should were responsible for some truly innovative hardware, not to mention a number other people from varying disciplines who made key contributions along the way. Anyway, suffice it to say that there were a lot of talented people involved in making Fred's dream a reality, and we all took great pride in our accomplishment. Indeed, as Earl Weltmer commented in the above-mentioned thread, it was quite a "work of art."
Timothy G. Moeller Senior Software Engineer NORAN Instruments Inc. 2551 W. Beltline Hwy. Middleton, WI 53562 tmoeller-at-noran.com
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We'd like to know what methods other labs are using to freeze down tissue for cryo-sectioning.
We're cooling down 2-methylbutane (iso-pentane) with dry ice and using the cooled liquid to freeze our tissue (which destined for immuno) in OCT. We don't have the facilities or the budget to use liquid nitrogen.
Are there methods that are better or use a less hazardous liquid? We want the tissue to freeze fairly quickly.
Thanks,
Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu
Center for the Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
by curly.cc.utexas.edu (8.8.5/8.8.5/cc-uts-1.20) with ESMTP id LAA05220 for {microscopy-at-sparc5.microscopy.com} ; Tue, 11 Aug 1998 11:47:50 -0500 (CDT)
Hi All, I'm tring to find a supplier of TEM grids that still offers custom photo- etching service of Au grids or screening. Has anyone used such services or could you point me in the right dirrection i.e. PolySciences, Ted Pella etc.
Alan, I'm not sure what type of analysis you are doing but if you are looking at sizing, most of the methods mentioned use a blow off or knock off. This would definitely skew your analysis. Getting a representative sampling of fine powders is a real challange. Finding a good method is very sample dependent. A good start for TiO2 would be a suspension of a powder sample in wetable water coupled with a 100% recovery on a small membrane filter. Russ
-----Original Message-----
Hi..having a problem with powder examination; have never before looked at powders (TiO2 in this case). Used conducting carbon adhesive pads to fix sample to stub but not much stayed on and although it coated ok most of sample came off whilst pumping out in the sample exchange chamber of our Hitachi S-800...How can i get round this..is there some sort fo fixative spray? Cheers Alan. Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
We are constructing some microscopy chambers and I have a vague recollection of reading some clever way to eliminate the meniscus effect so that ones gets an even buffer layer in a small chamber. Anyone have an idea whether I am dreaming or not? Thanks- Dave Knecht
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
} Linda- } we currently use this recipe: } Araldite 502 15ml } Eponate 12 (or equivalent) 25ml } DDSA 55ml } DMP-30 (catalyst)(1.5%) 1.25ml } } there are several mixtures (of varying hardness) in print, check any Hayat } or Glauert book on basic principals in EM. } as for reagents that are 10 years old (this stuff is cheap, your time is } not) buy new resins, plastics, DMP-30, etc. } -Mike } Linda & Mike -
A couple of days ago I suggested DMP-30 as the epoxy component that is most likely to go bad with long storage, but I didn't explain why. DMP-30 is inactivated by water, and it WILL hydrate if it isn't stored carefully. BDMA is an equally efficient accelerator, and is much less viscous than DMP-30;it can and should be used instead of DMP-30 in ANY epoxy recipe (Glauert, A.M. Accelerators for epoxy resins. Proc. R.M.S. 22:264,1987). The persistence of DMP-30 is a historical accident.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I'm tring to find a supplier of TEM grids that still offers custom photo- etching service of Au grids or screening. Has anyone used such services or could you point me in the right dirrection i.e. PolySciences, Ted Pella etc.
Dear Listers, If anyone knows of a source for a UV lamp - Cathodeon, TW6W black UV lamp for Leica's AFS in the states, would you let me know? Thanks in advance! Rosemary
Advanced Bioconcept produces a wide range of novel, high affinity, biologically active fluo-peptides for identification & characterisation of GPCR's, which can be very valuable to reaserchers in Confocal Microscopy.
They can be used to study peptide-receptor interactions in:
We have 31 established Fluo-peptides, and many are in development. If you would be kind enough to forward your contact details (fax number preferably), we would be very pleased to send you information about these products, such as protocols, a technical data sheets, and a product & price list. We can also outline our novel philosophy on custom syntheses if you wish.
Best regards,
Jon.
Jon Pinchuk ============================ Advanced Bioconcept Ltd. 1440 St. Catherine W., #424 Montreal, Qc, Canada, H3G1R8 http://www.bioconcept.com/ ============================ tel:514-874-5434 x22 fax:514-874-9077 mailto:jon-at-bioconcept.com ============================
Try Bulb Direct: 800-772-5267. Bulb Direct carries all kinds of bulbs under the sun. They may help you.
Regards,
In C. Kim, Ph.D. Ingen Laboratories, Inc. inkim-at-ingenlabs.com www.ingenlabs.com ---------------------------------------------------------- "Explorer of Molecular Hybridization"
-----Original Message-----
John N. Wright wrote: } } } Hi All, } I'm tring to find a supplier of TEM grids that still offers custom photo- } etching service of Au grids or screening. Has anyone used such services or } could you point me in the right dirrection. } } Thanks in Advance, } John N. Wright
-- Dear Mr. Wright,
We at Ladd Research would be happy to quote you. Please give us the mesh size requirements and the quantities and we will send you a quote or call the number below to discuss it.
Thanks,
John Arnott
LADD RESEARCH 13 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (FROM ANYWHERE) fAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.msa.microscopy.com/SM/LADD
The question of venting chambers and colums with dry nitrogen, using the boil-off gas from a liquid nitrogen container, has come up once or twice previously. As I mentioned then, there is a very inexpensive way to to this. What you do is to connect the vent tube of the liquid nitrogen container to the gas inlet of the chamber with a flexible tube (ordinary polyethylene tubing seems to work quite well, and is inexpensive and easy to work with). At a convenient spot along this tubing line insert a tee joint. Obtain a large inflatable-deflatable plastic beach ball and attach it to this tee joint with a length of flexible surgical rubber tubing. Using a razor blade or sharp scalpel, cut a clean slit about 100 mm long in this surgical rubber tubing. This slit then acts as a primative, but quite effective, pressure release valve. If the cut is clean, straight, and parallel to the axis of the tubing it will normally close tightly enough so that the nitrogen that boils off the storage container will accumulate in the beach ball. If the pressure in the ball rises significantly above that of the surrounding atmosphere, however, the slit will open slightly and allow the gas to escape. When the inlet valve to the vacuum chamber is opened, the slit will remain closed tightly enough so that no atmospheric gas can enter. The gas in the beach ball will then be driven into the chamber by only the pressure of the surrounding atmosphere so that overpressurization cannot occur. A beach ball that is about 2 ft in diameter will hold about 100 liters of gas, which is enough to fill most instruments several times, and of course the gas is being constantly replenished from the liquid nitrogen container.
If you are planning to use gas from a high pressure cylinder to fill EM chanbers, then you must be careful to ensure that you truly obtain oil-free gas in an oil-free tank. Ordinary compressed gases are often pumped with oil-sealed pumps and are contaminated with oil vapors from the pump, while ordinary tanks are usually contaminated with the oil vapors carried into them in this way.
These and related matters are discussed in some detail on pp. 63 - 65 of my book, 'Vacuum Methods in Electron Microscopoy' (see. http://www.cccbi.chester.pa.us/spi/catalog/books/book48.html, or http://www.bookshop.co.uk/portland/)
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-764-3321
I am attempting to cryosection starch-based foams, but they are collapsing in the TissueTek embedding medium because of the water content. Does anyone have ideas on making an embedding medium which is at least 95% anhydrous? I need the foams to keep their structure at least long enough for me to infitrate it in a vacuum(to get the medium into the air pockets) and then be frozen at about -20degC.
My "Correspondent's Report" from Inter/Micro 98 has just been posted to http://www.msa.microscopy.com/WWWviaEmail/EmailtoWWW.html . Please drop by and give it a read.
Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com
I would concur with a couple of other postings to your questions. The pointed filaments are not a good alternative. LaB6 should be the preferred cathode.
You don't give very specific details of your problem, but I see two possible problems. The first involves the manufacture of the LaB6 cathodes you use. You may be experiencing substantial amount of cathode drift after a few hours of use. This will result in a lower beam current, noisier image and a filament image that drifts to being eclipsed by apetures. In this case, I agree with another poster that Kimball Physics cathodes are quite exceptional. While you may associate this problem with the grid contamination, are you also re-centering your cathode when you clean the grid?
The other possibility is a vacuum leak. While you are able to attain and maintain a good overall vacuum, it is possible that there are vacuum leaks that direct incoming air to the cathode. Suspect the gun translation mechanism, if there is one, along with any vacuum standpipes or valves that open to the gun area. If the contamination is indeed the cause of your problems, an air leak is the most probable cause.
A properly designed and manufactured LaB6 cathode should not, on its own, produce grid contamination that would affect the imaging capability of the instrument. In fact, it would require a great deal of contamination to produce any visible effect on the imaging capability of an SEM in normally used magnification regions. If grid contamination is the actual cause of your problems, then you are either operating at the outside of the envelope for an SEM or the contamination is excessive because of vacuum systems leaks. On the other hand, you may be falsely attributing the cause to contamination where it really belongs to the poor design and manufacture of the cathodes you are using.
} Greetings, } } I was wondering whether anybody has any experience with pointed W } filaments in a XL-30. Ours currently has a normal W, although it is } design for LaB6 operation. The problem I have with LaB6 is poor } stability due to contamination build up on the Wehnelt (vac~2.5e-7 } mBar) and hence short operation time between cleaning (not filament } life time). The W is nice and stable and has been operating for } several months without a clean, however I need a bit more } brightness. } } Are there any advantages to using pointed filaments over normal? } What are the disadvantages? Are they worth the extra trouble? } } Keith. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr. K. Moulding. } } Materials Characterisation and Preparation Facility } Hong Kong University of Science and Technology, } Clear Water Bay, } Kowloon, } Hong Kong. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Thanks for the info. I was unfamiliar with the ADEM and since I service virtually every SEM made, I was quite curious when mention was made of it. I used to work with Pearson at ETEC. I had heard that he was helping with development of a new SEM, but never knew all the connections.
I'm sorry I've never had the opportunity to work with an ADEM. From your description, it sounds like they suffered in part from the common belief of the time that microcontrollers should be used where ever possible. Another example is ARL in the late eighties with their line of x-ray fluorescence spectrometers. While large numbers of microcontrollers can reduce the actual chip count of complex circuitry, the software development, debugging and basic understanding of the circuits being replaced has proven to limit their use in many cases.
I don't intend to disparage your vocation, rather I wish to point out that unrealistic expectations are often placed on it.
The development of a new SEM is a daunting and expensive task. The major manufacturers have it much easier, generally making small evolutionary changes. Producing a true revolutionary change requires money and perseverance. Those are things that few companies understand or can weather, particularily in today's corporate climate.
While the politics of Tracor's position may have lead to ADEM's demise, it may also have been its vision. Attempting to produce a revolutionary change from an evolutionary product may have ensured that the corporate expectations would be left wanting.
} Having just gotten back into the SEM community following a 12-year } hiatus, and while trying to get back up to speed by browsing through } the archives of this Microscopy list server, I read with extreme } interest the articles posted recently on the subject of the "ADEM1." } In that thread, Craig Theberge gave a partial answer from his own } personal experience to the question(s) that were raised, but I feel } compelled to fill in more of the historical details, completing the } picture from my own (albeit unique) perspective... } } ADEM was the internal code name for a major new secret project which } Fred Schamber headed up starting in 1985. (ADEM stood for "Advanced } Digital Electron Microscope", I believe, but was merely coined as a } code name and was not initially intended to be the actual name of } the product.) The development team was organized as a separate } company (later to be known as Tracor EBI) with it's own } multi-million-dollar budget and staff (consisting of only a handful } at the onset, handpicked from the Tracor-Northern R&D staff.) } Temporary facilities were rented off-site in the Middleton } Industrial Park (later dubbed the "Skunkworks") to house the entire } top-secret operation. We ended up staying there for the next } several years and even expanded the facility (renting adjacent bays) } to accommodate additional staff (adding on to engineering, but also } office personnel, stock, purchasing, drafting, etc.) Most people at } the main Tracor-Northern plant didn't know what was going on just } across town, and many didn't even know that the facility even } existed. It was being kept a secret for the reasons that Craig } mentioned - as a key microanalysis player at that time, we couldn't } afford to lose our alliances with existing SEM manufacturers until } we had perfected our own column. Fred was really going out on a } limb with this, putting the company's future at risk financially, } but felt he had the technical expertise to pull it off, and } corporate was willing to trust him because of his many previous } successes. After all, it would not have been an exaggeration to say } that Fred was largely responsible for building the success that TN } enjoyed by 1985, with a series of successful products dating all the } way back to the NS-880. } } Of course, in view of the unprecedented financial investment } involved, Fred was under considerable pressure to deliver a return } on investment in timely fashion and thereby prove that corporate's } confidence in him had not been misplaced. The core R&D personnel } involved at the inception were also quite dedicated to the success } of the project, and so you can bet that for the next couple years, } there were plenty of late hours and weekends devoted by all. Most } of these have gone their separate ways now, but they included Mike } Krummey (electronics engineering), Jorgen Rasmussen (mechanical } engineering) and myself. We also had two primary consultants on the } project: Dave Pearson (an experienced Etec serviceman, who brought } in an old junker Etec scope which he kept running for us to use as a } test bed for proof-of-concept on several hardware design issues), } and Rich Lee! Yes, that's the same Rich (R.J.) Lee whom Fred later } joined to produce the Personal SEM. A few other individuals were } brought in from the outside as the need for certain skills became } apparent - Ken Spielman, for example (who is still at Noran, by the } way), was a skilled machinist hired in from the UW to hand-machine } all the various sundry precision metal parts needed in the } construction the first prototype. And of course there were } technicians, draftsmen, purchasing and office support personnel, } etc., added as time went on (and even marketing, sales and service } people still later.) But it was Fred who spearheaded and } masterminded the whole operation. It was his dream initially, his } brilliance as a physicist which was crucial in solving several key } problems along the way, his resourcefulness which kept the project } on budget, and it was his drive and determination which kept the } project moving ahead. } } Fred had his hand in nearly every aspect of the development, but } soon was spending the majority of his time focusing (pun intended) } on the design of the lenses, which involved sophisticated } electromagnetic field computations as well as careful selection of } appropriate alloy materials for the pole pieces, etc. But despite } the complexities involved in that, and with all due respect to Fred, } those were among the more conventional aspects of the ADEM. What } made ADEM truly unique was its huge chamber, sophisticated and } fully-articulated sample stage, along with complete integration of } all subsystems and digital control of each utilizing distributed } microprocessor control. It was my pleasure to develop software and } firmware for several parts of the latter, including the entire } six-axes stage controller (consisting of microprocessors to drive } each of the three translational and three rotational axes, and a } "master" processor to coordinate the motions of all six axes.) In } fact, microprocessors drove every subsystem of the ADEM, including } not only the stage, but also the scan generator, vacuum system, EDS } front end, etc., even the water valve! Dear reader, please indulge } me as I proudly mention that I was responsible for the embedded } software on nearly all of these subsystems, and personally wrote the } code for most of them. But the whole instrument was tied together } by a central processor which also performed the primary imaging } functions; and the software for that part of the system was } developed by a team of several other talented programmers. Of } course, we were working with several bright electronics and } mechanical engineers who should were responsible for some truly } innovative hardware, not to mention a number other people from } varying disciplines who made key contributions along the way. } Anyway, suffice it to say that there were a lot of talented people } involved in making Fred's dream a reality, and we all took great } pride in our accomplishment. Indeed, as Earl Weltmer commented in } the above-mentioned thread, it was quite a "work of art." } } Timothy G. Moeller } Senior Software Engineer } NORAN Instruments Inc. } 2551 W. Beltline Hwy. } Middleton, WI 53562 } tmoeller-at-noran.com } } Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
Does anyone have a good method for cleaning powder metal samples that have been emmersed in oil? Specifically, these parts are saturated with transmission fluid and it is extremely difficult to examine them in the SEM without some seeping of the fluid from the interior of the part. Past attempts have included vacuum, a wide range of solvents, sonication as well as heating to burn off any remaining fluid. Any info is greatly appreciated.
Dear all, in our TEM-Lab runs a discussion about the use of uranylacetate in routi= ne secion-staining because of its radioactivity and the possible risk of ca= ncer. We are wondered about the relative 'light' safety instructions for handl= ing and disposal uranylsolutions compared to that for C-Isotopes used in special enzymat= ic reactions. 1.Is this risk negligible , if not, what intern al safety instructions exi= sts in other labs or is literature about the risk of uranylacetat in biological labs available? 2.Has anybody experience with alternative (non-radioactive) counter stains= ? Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=E4tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
Thank you for the kudos. And, no offense taken regarding the fact that we went overboard with the use of microprocessors in the design of the ADEM - in fact, I agree with you. Indeed, that is one of the reasons for it's short life (after all, the original poster which prompted my little expose' was trying to dispose of two "old" ADEMs already, not even 10 years old!) They are just too complex to maintain &/or modify. And, as proud as I am of my embedded code, I must admit that the fact that it is entirely proprietary and not user-programmable makes it extremely inflexible.
You are certainly correct about the development of a new SEM being "a daunting and expensive task." The ADEM development team was all too well aware of that, even from the start. And we had that truth painfully impressed upon us ever more so each day as we struggled with trying to get it done. It would be really nice to hear from Fred (Dr. Frederick H. Schamber, now with RJ Lee Instruments, as I mentioned) to fill in even more of the background. He bore the brunt of many of the criticisms which you have voiced concerning the decision to develop the ADEM, and I'm sure would be able to offer further insights into the rationale for doing so. If nothing else, I believe he would mention that there was some motivation for creating an SEM entirely made in the U.S.A., at a time when the American work ethic was being challenged by the Japanese, and blamed for the growing trade deficit. At any rate, I don't think anyone could really fault him for the decision; at least his motives were pure, even if he lacked some business savvy. And he was certainly up to the task from a technical standpoint; after all, if anybody could pull it off, Fred could - he's a uniquely talented, extremely intelligent and exceptionally energetic guy, for whom I have the utmost respect and admiration. The SEM community is blessed to still have him in their midst, even if it's just working on " evolutionary" SEMs.
Another bit of historical information (completely unrelated) that I just remembered... At the same time we were developing the ADEM, the birth of our first CONFOCAL microscope took place. Dr. D. O. Landon got a few of us involved at the "Skunkworks" off in a corner, playing on an optical table with lasers, motion controllers and other cool "toys", actually acquiring high-resolution images of biological specimens. I even wrote some TN-5500 software (in FLEXTRAN) in my spare time to control and display images from that very first confocal microscope. But of course that seemed like just a fun diversion; we didn't regard this as an important development at the time. Oddly enough, however, confocal microscopy remains a significant part of the core business here at NORAN, second only to the microanalysis instrumentation products, and confocal are the only microscopes we manufacture at all anymore. Kind of shows the value of always keeping something on the "back burner", I guess.
Timothy G. Moeller Senior Software Engineer NORAN Instruments Inc. 2551 W. Beltline Hwy. Middleton, WI 53562 (608) 836-4119 tmoeller-at-noran.com
----------
Thanks for the info. I was unfamiliar with the ADEM and since I service virtually every SEM made, I was quite curious when mention was made of it. I used to work with Pearson at ETEC. I had heard that he was helping with development of a new SEM, but never knew all the connections.
I'm sorry I've never had the opportunity to work with an ADEM. From your description, it sounds like they suffered in part from the common belief of the time that microcontrollers should be used where ever possible. Another example is ARL in the late eighties with their line of x-ray fluorescence spectrometers. While large numbers of microcontrollers can reduce the actual chip count of complex circuitry, the software development, debugging and basic understanding of the circuits being replaced has proven to limit their use in many cases.
I don't intend to disparage your vocation, rather I wish to point out that unrealistic expectations are often placed on it.
The development of a new SEM is a daunting and expensive task. The major manufacturers have it much easier, generally making small evolutionary changes. Producing a true revolutionary change requires money and perseverance. Those are things that few companies understand or can weather, particularily in today's corporate climate.
While the politics of Tracor's position may have lead to ADEM's demise, it may also have been its vision. Attempting to produce a revolutionary change from an evolutionary product may have ensured that the corporate expectations would be left wanting.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-mcs.net WWW: http://www.mcs.net/~ars Analytical instrument maintenance services
I am going to be transiting in Miami on my way to Cancun and have been told that I would need a VISA for these few minutes in the United States. Is that really true? Kristof Kovacs
************************************************************************* Dr. Kristof KOVACS President, Hungarian Society for Microscopy Associate Professor University of Veszprem Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684 Fax: +36-(88)-328-643 *************************************************************************
We routinely use the iso-pentane and dry ice method with good results on skin biopsies. We also use ethanol and dry ice with the same results. You just have to be very careful not to let the ethanol come in to contact with the OCT. It will make it rubbery and can't be cut.
The other issue is if you need to fix the tissue you need to cryo-protect it after fixation before you embed and freeze it, or it will shread as it thaws. We use Zambonies or paraformaldehyde, then after rinsing in PBS, cryoprotect in 8.5% sucrose in PBS for two hours then embed in OCT and freeze.
Bob Derm Imaging Center U of W
On 11 Aug 1998, Jaci Lett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We'd like to know what methods other labs are using to freeze down tissue for } cryo-sectioning. } } We're cooling down 2-methylbutane (iso-pentane) with dry ice and using the } cooled liquid to freeze our tissue (which destined for immuno) in OCT. We don't } have the facilities or the budget to use liquid nitrogen. } } Are there methods that are better or use a less hazardous liquid? We want the } tissue to freeze fairly quickly. } } Thanks, } } Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu } } Center for the Biology of Hearing and Deafness } Central Institute for the Deaf } 818 S. Euclid Ave. } St. Louis, MO 63110 }
A lady replied on my query regarding MEIJI microscopes.
I would like to say thank you for taking the effort to reply and I am sorry but I have mistakenly deleted your message. Could you perhaps E-mail me again...I would like to discuss perhaps the optics in more details. I am of the opinion at the moment that the optics are not good enough to waraant equiping it with a digital camera since there are already enough problems with image analysis from a good image let alone a poor one.
And if anyone out there could make recommendations on optical microscopes. For example, I know Zeiss is supposed to be the best, with Leica and Nikon following. I want to know more than just the brandname, but I want information on the exact optical configuration and why it is better or worse than another, because suppliers seem to be too secretive, so I am asking USERS for their expert opinion based on sound experience.
I would appreciate very much any avalaible information or at least sources where I could find this kind of information.
Cherio, Quirina
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
Jaclynn, My opinion after freezing tons of tissue for four years is that 2-methylbutane cooled in dry ice is the way to go- quick,simple,cheap. I made sure the 2-mb was cooled enough by putting in a few small pieces of dry ice until the vigorous bubbling stopped or slowed almost completely (at which the temperatures are nearly equal). I also made sure that any tools were cooled beforehand. Lastly, when the tissue is plunged quickly into the cooled 2-mb, move it around gently to keep it in contact with cooled fluid. Have you considered cryoprotectants? Are you experiencing problems? "Practical Methods in Electron Microscopy, vol 13, Sectioning and Cryosectioning for Electron Microscopy," is a good reference.
-------------------------------------------------------- Winston W Wiggins, Supr. 8/12/98 10:49:56 AM CRC-Electron Microscopy Lab. Ofc:704/355-1267 Carolinas Medical Center Fax:704/355-7648 P.O. Box 32861 Lab:704/355-7220 Charlotte,NC 28232-2861 USA Eml:wwiggins-at-carolinas.org --------------------------------------------------------
We are trying to do immunostaining of newt tissue, which, while alive, was expressing GFP. Cryosectioning is an option. I heard that prolonged fixation in paraformaldehyde will quench the GFP signal - is that true? The decision to make is whether to prefix the tissue for longer period of time and risk losing the signal or quick freeze, section and then fix for shorter period of time. Also, what would be an optimal fixative for GFP retention?
Sharing any thoughts or direct experience in this area would be highly appreciated.
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
The program and preliminary poster for the joint NYSEM and AIF all day symposium at the Albert Einstein College of Medicine on Sept. 10 is posted at http://www.ca.aecom.yu.edu/aif/directions.htm
-------------------------------------------- Michael Cammer email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 http://www.ca.aecom.yu.edu/aif/ --------------------------------------------
We are looking for two used research quality inverted microscopes, e.g., Zeiss IM35, Nikon Diaphot 200 or later, or Olympus I50 or 70, for patch clamp recording purposes. If anyone has ones available please email: yamoahen-at-email.uc.edu, or call Ebenezer Yamoah at 513-558-4909.
I have been trying to silver enhance 1nm gold label that is embedded in LR White sections. After prolonged (90min) soaking in BBI sliver enhancing solution (SEK15), the surface particles are very large, but the enhancement is not the penetrating the plasticand getting to the gold trapped in the section. Does anyone have a method for permualizing LR White, so that enhancing solution penetration will occur?
TIA
William R.McManus Supervisor Electron Microscope Facility Department of Biology Utah State University Logan UT 84322-5305
(1.38.193.4/16.2) id AA20297; Thu, 13 Aug 1998 07:50:13 -0400 Received: by DA_EXC1.sylvania.com with Internet Mail Service (5.5.2232.9) id {Q5KXC88S} ; Thu, 13 Aug 1998 07:12:28 -0400
I'm looking for an independent failure analysis lab in the Southwestern US. The lab should be capable of performing microanalytical techniques on glasses, ceramics, metals, plastics and composites. Commercial and academic labs are acceptable. Please respond to me directly.
Thanks,
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
I have been playing around with sovelnts (diff pump stack revival) Carbon tertracloride CCl4 fumes are brilliant. It boils at 76.54 deg Centegrade. Be sure that you work in a fumehood! Is used as a degraser at Cr and Ni coating facilitys. } } Does anyone have a good method for cleaning powder metal samples that have } been emmersed in oil? Specifically, these parts are saturated with } transmission fluid and it is extremely difficult to examine them in the SEM } without some seeping of the fluid from the interior of the part. Past } attempts have included vacuum, a wide range of solvents, sonication as well } as heating to burn off any remaining fluid. Any info is greatly } appreciated. } } Wayne England } wengland-at-ortech.on.ca }
Mr. S H Coetzee Tell: (011) 716 2419 Electron Microscope Unit Fax: (011) 339 3407 Private bag X3 E-mail: Stephan-at-gecko.biol.wits.ac.za Wits Johannesburg 2050
Attached is the TOC for the journal MICRON for August 1998
CONTENTS LIST Date 13-JUL-1998 Page 1
Journal : Micron Volume/issue : 29/4 Year : 1998 ISSN : 0968-4328 Cover Date : August 1998
pp. iv-iv Editorial
pp. 251-259 PII S0968-4328(97)00062-0 JMIC 232 Ed. office no. E53 Microstructure of polarized electrochemical vapor deposition (PEVD) products EZ Tang, DG Ivey, TH Etsell
pp. 261-265 PII S0968-4328(97)00063-2 JMIC 233 The significance of locating and filling the canal isthmus in multiple root canal systems. A scanning electron microscopy study of the mesiobuccal root of maxillary first permanent molars DC Yu, A Tam, MH Chen
pp. 267-280 PII S0968-4328(97)00064-4 JMIC 234 Prosobranch parasperm: sterile germ cells that promote paternity? J Buckland-Nicks
pp. 281-287 PII S0968-4328(97)00057-7 JMIC 227 Ed. office no. E 52 Microstructural characterization of Au/Sn solder for packaging in optoelectronic applications DG Ivey
pp. 289-292 PII S0968-4328(97)00053-X JMIC 223 Ed. office no. E56 Events at the university of Alberta and the university of Toronto, leading to the first North-American electron microscope AF Prebus
pp. 293-296 PII S0968-4328(98)00013-4 JMIC 257 Modification of unicryl composition for rapid polymerisation at low temperature without alteration of immunocytochemical sensitivity P Gounon, J-P Rolland
pp. 297-307 PII S0968-4328(98)00011-0 JMIC 255 Optimization of phosporous localization by EFTEM of nucleic acid containing structures C Quintana, S Marco, N Bonnet, C Risco, ML Gutierrez, A Guerrero, JL Carrascosa
pp. 309-328 PII S0968-4328(98)00015-8 JMIC 259 Aspects of the structure and assembly of desmosomes IDJ Burdett
Dear Judy, Long fixation may reduce GFP signal. Paraformaldehyde fix of 30 minutes should be sufficient for most systems. However. You will lose all GFP signal if it is a free GFP in the cytosol or nucleus upon permeabilization due to extraction. You do not mention whether this is a GFP fusion protein. GFP- fusion proteins that a integral parts of membranes or structures will remain.
Joseph Goodhouse Confocal / EM Core Lab Manager Department of Molecular Biology Princeton University jgoodhose-at-molbio.princeton.edu 609-258-5432 Info and Images at http://www.molbio.princeton.edu/confocal/CF-EM-HOME.html
} -----Original Message----- } From: Judy Trogadis [SMTP:judy-at-playfair.utoronto.ca] } Sent: Wednesday, August 12, 1998 11:26 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: GFP immuno } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } We are trying to do immunostaining of newt tissue, which, while alive, } was } expressing GFP. Cryosectioning is an option. I heard that prolonged } fixation } in paraformaldehyde will quench the GFP signal - is that true? The } decision } to make is whether to prefix the tissue for longer period of time and } risk } losing the signal or quick freeze, section and then fix for shorter } period } of time. Also, what would be an optimal fixative for GFP retention? } } Sharing any thoughts or direct experience in this area would be highly } } appreciated. } } } Judy Trogadis } Eye Research Institute and } University of Toronto } Toronto Hospital, Western Div. } 399 Bathurst St. } Toronto, Canada M5T 2S8 } } phone: 416-603-5088 } Fax: 416-603-5126 } email: judy-at-playfair.utoronto.ca }
It seems most of the avid writers are conferencing, so here=20 is my contribution: This forum has dealt with UA safety several times before.=20 It is, however, very time consuming to go through a year of=20 the archives, so here are is my summary/opinion.
1 All elements above lead in the periodic table are=20 radioactive. 2 Carbon, phosphorus and other isotopes are readily=20 absorbed and incorporated in tissues. That makes these=20 "biological isotopes" more dangerous. Uranyl Acetate is water soluble and is not stored in the=20 body. 3 Whereas in mining, the insoluble Uranium particles are=20 lodged in the lung and emit radon gas for many years and=20 this makes insoluble U compounds a much greater=20 radiological hazard. 4 UAs' chemical toxicity is greater than its radiation=20 hazard - just don't ingest that stuff, it's a powerful=20 kidney poison, but happily not cumulative. 5 In general terms, the higher an element on the periodic=20 table, the more intense the electron "staining". A good=20 argument for lead, being the last non-radioactive element,=20 except that it a cumulative toxin, but we use it with care=20 (I trust). Actually I have seen lead pigments used in an=20 industrial setting (36 yrs ago), truly hair raising by=20 today's standards, but it was known then that lead was a=20 cumulative poison. 6 The (Edelmetalle) Au, Pd, Pt, Ir would seem attractive=20 alternatives for electron staining, but they are=20 non-reactive and only useful as markers or in evaporation=20 techniques. 7 Dense, high molecular number compounds have some staining=20 effects, for instance Sudan Black to show lipids, but these=20 stains are not nearly as effective as are the high atomic=20 number elements. 8 I am worried why so many people are worried. You would=20 have cause if you were dealing with the 200 litre drum=20 quantities of U "yellowcake", which has very similar=20 toxicity and radioactive attributes. 9 Consider: You are in a laboratory with fumehood and=20 gloves available, you are rather more knowledgeable about=20 these matters then armies of industrial workers and you are=20 using tiny quantities. 10 On balance I would suggest that it is rather more=20 dangerous to fly at high altitudes because of the gamma=20 radiation and it's a terrible thing to eat steak, because=20 of the high fat content and any scorched parts are=20 carcinogenic.
If you handle in a laboratory setting UA prudently, it is=20 in my opinion one of the less hazardous encounters in life.=20 I suggest that even a walk in the Teutoburger Wald, not to=20 mention urban Bielefeld is not without its dangers. Cheers Jim Darley ProSciTech Microscopy=20 PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au=20 *****
On Wednesday, 12 August 1998 23:25, B. Laube=20 [SMTP:B.Laube-at-biologie.uni-bielefeld.de] wrote: } Dear all, } in our TEM-Lab runs a discussion about the use of } uranylacetate in routine } secion-staining because of its radioactivity and the } possible risk of cancer. } We are wondered about the relative 'light' safety } instructions for handling and disposal } uranylsolutions compared to that for C-Isotopes used in } special enzymatic reactions. } 1.Is this risk negligible , if not, what intern al safety } instructions exists in other labs or is } literature about the risk of uranylacetat in biological } labs available? } 2.Has anybody experience with alternative (non- } radioactive) counter stains? } Bernward Laube } University of Bielefeld } Faculty of Biology } Department Plant Morphology and Cell Ultrastructure } Universit=E4tsstrasse 25 } Germany 33615 Bielefeld } phone: 0521 1065592 } fax: 0521 1066039 } e-mail: b.laube-at-biologie.uni-bielefeld.de } http://www.uni- } bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
hello....anybody know an easy way i.e. via a macro or similar to get a conversion from the ASTM G grain size number to the actual value in real dimensions?..is it different for the various ASTM grain size measurement types that are available ,i.e horizontal, vertical, diagonal and concentric circles. The image analysis package we have is Image Pro Plus with Materials Pro from Media Cybernetics (USA) which gives us the grain size in the ASTM G unit. I have info from the ASTM standards E1382 and E112 concerning this but there is a lot of info in there I'm not sure about. cheers. alan. ----- Dr Alan Templeton Centre for Physical Electronics and Materials SEEIE South Bank University 103 Borough Road, London SE1 0AA TEL 44 171 815 7521 /7571 FAX 44 171 815 7599 email templea-at-sbu.ac.uk
We use sodium m-periodate saturated, for about 10-20 min to etch away some of the LRwhite but I have never tried this on prelabelled samples. I would think it might strip away the gold.
bob Derm Imaging Center U of W
On Wed, 12 Aug 1998 billemac-at-cc.usu.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have been trying to silver enhance 1nm gold label that is embedded in LR } White sections. After prolonged (90min) soaking in BBI sliver enhancing } solution (SEK15), the surface particles are very large, but the enhancement } is not the penetrating the plasticand getting to the gold trapped in the } section. Does anyone have a method for permualizing LR White, so that } enhancing solution penetration will occur? } } TIA } } } William R.McManus } Supervisor } Electron Microscope Facility } Department of Biology } Utah State University } Logan UT 84322-5305 } } billEMac-at-cc.usu.edu } Ph 435-797-1920 } Fax 435-797-1575 } } }
} in our TEM-Lab runs a discussion about the use of uranylacetate in routine } secion-staining because of its radioactivity and the possible risk of cancer. } We are wondered about the relative 'light' safety instructions for handling } and disposal uranylsolutions compared to that for C-Isotopes used in spe- } cial enzymatic reactions. } 1.Is this risk negligible ,
A qualified yes. The amount of activity is very low, and U is an alpha-particle emitter. The range of the alphas is less than the thickness of the dead layer of skin, so UAc is not a radiation hazard if one gets it on ones hands, etc. However, alpha emitters can be extremely hazardous if they are inhaled or ingested, so precautions should be taken so that small droplets are not produced, and always wash your hands after using UAc.
} if not, what intern al safety instructions exists in other labs or is } literature about the risk of uranylacetat in biological labs available?
Your safety office should have info on internal procedures; these can vary from place to place. In the US we can get a materials safety data sheet (MSDS) for any substance, and this will give info about hazards, re- strictions for transport and use, etc. I'm sure any of the local EM sup- pliers can get this for you. Yours, Bill Tivol
Kristof Kovacs wrote: ================================================ I am going to be transiting in Miami on my way to Cancun and have been told that I would need a VISA for these few minutes in the United States. Is that really true? ================================================= There seems to be a lot of ambiguity about this and if you call the same airline three times, you can get three different opinions on this. I can understand how there could have been some confusion on this.
We are talking about someone travelling to Cancun, transiting some point in the USA en route to Cancun, and they would be travelling on a passport for which a visa for travel to the USA would normally be required. Anyone else has no worries whatsoever.
I called United Airlines three times and received three different answers. I called Lufthansa twice and got still other answers. But this is what seems to be the correct situation:
If a passenger (as described above) arrives in the USA as a transit passenger, if they are holding a confirmed onward ticket for entry into a third country, you are exempted from the visa requirement and you do not need a visa. There is one exception to this and that is, if the transit time is more than eight hours. Then you lose your eligibility for the exemption and you would need a visa (according to "Linda" at LH). Someone "overnighting" at their transit point could fall into this category.
This should not be a cause for concern or worry. If you do have any such concerns, contact the airline bringing you to the USA point of entry, and make sure that in your record they have "documented the record" as to what passport you are travelling on and that you won't need a visa for the purposes of transiting the USA. That could save you any kind of hold up or inconvenience when you check in for your first flight.
Linda told me one other interesting thing: She said that sometimes people travel to Cancun and like it (literally) so much, they stay more than 90 days and then they run into a visa requirement of Mexico!
Hope this clarifies things.
Chuck
PS: Remember I am not an expert on this, that is why if you have any further concerns, contact your carrier to the USA for the "last word" on this. And be sure they document what they tell you in your record!
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} Date: Wed, 12 Aug 1998 11:26:21 -0400 (EDT) } From: Judy Trogadis {judy-at-playfair.utoronto.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: GFP immuno } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are trying to do immunostaining of newt tissue, which, while alive, was } expressing GFP. Cryosectioning is an option. I heard that prolonged fixation } in paraformaldehyde will quench the GFP signal - is that true? The decision } to make is whether to prefix the tissue for longer period of time and risk } losing the signal or quick freeze, section and then fix for shorter period } of time. Also, what would be an optimal fixative for GFP retention? } } Sharing any thoughts or direct experience in this area would be highly } appreciated. } } } Judy Trogadis } Eye Research Institute and } University of Toronto } Toronto Hospital, Western Div. } 399 Bathurst St. } Toronto, Canada M5T 2S8 } } phone: 416-603-5088 } Fax: 416-603-5126 } email: judy-at-playfair.utoronto.ca
We do not have trouble with p-form fixed GPF tissue. What do you call "prolonged."} } If you want to look at ultrastructure, you CANNOT freeze and then fix; you must fix first. I need to know more about what you want to do before commenting on methods. S}
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We have got lots of historical Journals available for those, who are interested in getting a set or filling some shortages in his or his departements library. Please contact me, I will forward your inquiry to Prof. Hildebrand. Please be aware that the delivery charge for shipment has to be payed by the recipient.
Please find attached the counts of the existing volumes.
Can anyone tell me for certain how much BDMA is equivalent(volume to volume) to DMP-30? There are very confusing substitution rules to be found in the literature. Can anyone guarantee the same result if we suddenly switch to BDMA? We really don not have time to run tests. DMP-30 is problematic with its capability of binding water. We use it only for a short time, then discard the rest. But, one never knows how long the reagent has been sitting on a shelf before purchase! Thanks, Hildy Crowley {hcrowley-at-du.edu}
Dear colleagues, I would like to find out if there is a non- fluorescent dye that can be used on fluorescently labeled sections (to identify structures) without compromising the integrity of the fluorescence? Thank you, Lilith --------------------------------------- Lilith Ohannessian-Barry NRC, IBS, Ottawa, Ont. K1A 0R6 Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
} Hi, } } Can anyone tell me for certain how much BDMA is equivalent(volume to } volume) to DMP-30? There are very confusing substitution rules to be } found in the literature. Can anyone guarantee the same result if we } suddenly switch to BDMA? We really don not have time to run tests. } DMP-30 is problematic with its capability of binding water. We use } it only for a short time, then discard the rest. But, one never knows how long the reagent has been sitting on a shelf before purchase! } Thanks, } Hildy Crowley } {hcrowley-at-du.edu}
Hildy:
I have used BDMA in my "Epon" mixes for about 2 years, absolutely no problems. I use 2.5% to 3% BDMA as recommended by Electron Microcopy Sciences in the kit they supply. I do everything by weight. I replace the BDMA 6-8 months after opening, just to be safe and it is stored at room temp.
Geoff
-- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Would any interested Third Party Maintenance organizations drop me an e-mail including their location, area of expertise, main contact, etc. I wish to form a clearinghouse for all TPMs to share technical information, customer referral, etc.
I have been in business as an SEM third party maintenance organization in Southern California for over 15 years. Quite often I am asked to recommend someone or repair equipment outside our geographical area.
We have everything to gain. The ultimate winner is the customer.
I have obtained a Vickers Shearing Eyepiece. It fits in place of an eyepiece in a microscope and apparently through a set of prisms and colored filters it forms two images (one red and one green) that can be separated by means of a micrometer dial in the eyepiece.
Does anyone know what this was used for. My best guess is that it may have been used for measuring particle sizes by "separating" the two images from each other and then reading the micrometer scale.
Dear all, I have had a student from another department come to see me needing some help for his project. He needs a "method for the preparation and analytical procedure to determine algal metal uptake on cell surface and in the various subcellular regions". If anyone has any ideas I would appreciate it very much. TIA Joan Clark
Would any interested Third Party Maintenance organizations drop me an e-mail including their location, area of expertise, main contact, etc. I
wish to form a clearinghouse for all TPMs to share technical information, customer referral, etc.
I have been in business as an SEM third party maintenance organization in Southern California for over 15 years. Quite often I am asked to recommend someone or repair equipment outside our geographical area.
We have everything to gain. The ultimate winner is the customer.
} There seems to be a lot of ambiguity about this and if you call the same } airline three times, you can get three different opinions on this. I can } understand how there could have been some confusion on this.
This is true, mainly because there are different rules for nationals of different countries. When this question arose a couple of days ago my travel agent provided me with a copy of the regulations (about 500 words) applying only to persons travelling in transit through the US!!
If anyone is interested I can look up details from this document, otherwise this information is available from any US Embassy or Consulate or most airlines and travel agents.
Robin
} We are talking about someone travelling to Cancun, transiting some point in } the USA en route to Cancun, and they would be travelling on a passport for } which a visa for travel to the USA would normally be required. Anyone else } has no worries whatsoever. } } I called United Airlines three times and received three different answers. } I called Lufthansa twice and got still other answers. But this is what } seems to be the correct situation: } } If a passenger (as described above) arrives in the USA as a transit } passenger, if they are holding a confirmed onward ticket for entry into a } third country, you are exempted from the visa requirement and you do not } need a visa. There is one exception to this and that is, if the transit } time is more than eight hours. Then you lose your eligibility for the } exemption and you would need a visa (according to "Linda" at LH). Someone } "overnighting" at their transit point could fall into this category. } } This should not be a cause for concern or worry. If you do have any such } concerns, contact the airline bringing you to the USA point of entry, and } make sure that in your record they have "documented the record" as to what } passport you are travelling on and that you won't need a visa for the } purposes of transiting the USA. That could save you any kind of hold up or } inconvenience when you check in for your first flight. } } Linda told me one other interesting thing: She said that sometimes people } travel to Cancun and like it (literally) so much, they stay more than 90 } days and then they run into a visa requirement of Mexico! } } Hope this clarifies things. } } Chuck } } PS: Remember I am not an expert on this, that is why if you have any } further concerns, contact your carrier to the USA for the "last word" on } this. And be sure they document what they tell you in your record! } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== }
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/affiliates/emu/em.htm
I have had experience with animal tissues. Presumably, you are contemplating x-ray microanalysis. I would avoid all chemical fixatives !! They cause leaching of soluble ions etc. The specimen needs to be cryofixed in some way (even rapid freezing on a cold copper plate cooled by liquid nitrogen). Then, freeze dried (even by putting them in a vacuum coating unit on a cold copper plate (on plastic insulation to prevent fast warm-up), and allowing to pump for a couple of days. Then sections? Infiltrate in liquid resin under vacuum to assist penetration, take out, change and cure as normal. Then cut 0.5 micron sections on a DRY glass knife, put onto film-coated grids (I use colloidion for ease of preparing them) which may be pre-coated with carbon. Then carbon coat (against charging in STEM). The grid material may be copper, aluminium, titanium, nickel - they may all fluoresce and give false results so be careful!
Hope this gives some idea - it can be done very usefully this way but the ultrastructure will probably be ***p! Freezing damage will be quite extensive but the goodies will all be in there. With freeze drying, some of the crystal damage will be masked because I believe some membranes will 'relax' before being caught in position by the hardening of the resin.
Another approach is to freeze (as above) and then freeze substitute for about 10 days in pure acetone at -80. This can be done simply by suspending the specimens in a container above liquid nitrogen. I use a one-third-full 25 litre dewar with the specimens about 2 inches from the top. You must use a thermocouple or somehow determine this temp. I have a wire tc to a digital thermometer which sits in the base of a brass gauze basket containing the specimens. Then bring to room temperature and infiltrate with resin etc. This method was really meant fr low temperature embedding (which I don't have).
Hope this helps - Keith Ryan Plymouth Marine Lab., UK
} Does anyone know what this was used for. My best guess is that it may have } been used for measuring particle sizes by "separating" the two images from } each other and then reading the micrometer scale.
Exactly. As I remember, you displace one image until it is tangential to the other and read off the diameter (displacement).
I am a designer in Dallas and am doing a poster for a trade show for one of out clients and need to purchase a color scan of dust, spores, mites etc.. to show what type of organisms live in the air we breathe contact me via e-mail and we can discuss cost and availability. my address is jharskjold-at-gpcreative.com.
Hi All: I am looking for a sure-fire method to remove the "speckled" appearance in silicon HREM images. I've heard of etchants and other methods to remove this artifact, but I am unaware of any specific sure-fire recipes/methods. Any contributions would be greatly appreciated. Thanks in advance.
Regards,
Michael Coviello UT Arlington Materials Science Arlington, TX 817 272-5496
Joan, I have done extensive EM on the uptake of metals by different = species of algae and bacteria under experimentally controlled = conditions.. In these cases the algae were incubated with different = concentrations of selected heavy metals. Under these conditions the = metals were adsorbed/ppt to the cell walls of living algae or bacteria = and were clearly visible as discrete particles doing conventional TEM = without contrast enhancement. Under these controlled conditions in which = the metal compounds are added to the media it may not be necessary to do = edx unless you want to do semiquanitative analysis although the = differences will be clearly visible. When the organisms are dying or = already dead, uptake is intracellular/internal. Prepare the samples = with or without osmium postfixation. Postfixation in somecases augments = the visibility depending on the metals. This is a good start and will = reveal a lot of info. It can get complicated and expensive if you want = to go farther with it. Obviously, uptake of metals which are unbound will be loss by this = procedure in which case quenching in a LN2/propane or isopentane = followed by freezesubstitution or freeze drying and embedding or = cryosectioning, etc. is necessary. Hank Adams IMC Cell Biology Baylor College of Medicine
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} Joan, I have done extensive EM on = the uptake of=20 metals by different species of algae and bacteria under experimentally=20 controlled conditions.. In these cases the algae were incubated with = different=20 concentrations of selected heavy metals. Under these conditions = the metals=20 were adsorbed/ppt to the cell walls of living algae or bacteria and were = clearly=20 visible as discrete particles doing conventional TEM without contrast=20 enhancement. Under these controlled conditions in which the metal = compounds are=20 added to the media it may not be necessary to do edx unless you want to = do=20 semiquanitative analysis although the differences will be clearly = visible.=20 {/FONT} {FONT color=3D#000000 size=3D2} When the organisms are dying or = already=20 dead, uptake is intracellular/internal. Prepare the samples = with or=20 without osmium postfixation. {/FONT} {FONT size=3D2} Postfixation in = somecases=20 augments the visibility depending on the metals. This is a good start = and will=20 reveal a lot of info. It can get complicated and expensive if you = want to=20 go farther with it. {/FONT} {/DIV} {DIV} {FONT size=3D2} Obviously, uptake of metals which are unbound = will be=20 loss by this procedure in which case quenching in a LN2/propane or = isopentane=20 followed by freezesubstitution or freeze drying and embedding or = cryosectioning,=20 etc. is necessary. {/FONT} {/DIV} {DIV} {FONT size=3D2} Hank Adams {/FONT} {/DIV} {DIV} {FONT size=3D2} IMC Cell Biology {/FONT} {/DIV} {DIV} {FONT size=3D2} Baylor College of = Medicine {/FONT} {/DIV} {/BODY} {/HTML}
I have located in the archives instructions/schematics for a Kinney SC 3 evaporator. The machine has long since been cannibalized. Anyone wanting this material is welcome to it--otherwise it goes into "file 13" shortly.
Just email with address if interested.
S
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
The Vickers Shearing Eyepieces were well known for their high accuracy and precision in measuring particle sizes. I had the privilege of using them while on an RMS short course in the UK 20 years ago. You are right about the image separation. While I really need to dig out the instructions to be absolutely sure of they work, I remember the following: That you superimposed the images and either set the vernier to zero or read the vernier setting. Next, you rotated the drum until the two images were separate but just touching and read again. It may be that you need to divide the results by 2. Try it with something of known diameter to test. I will rummage through the extensive MME archives and see if I can find more complete instructions. In the meantime, enjoy your new found treasure!
Best regards,
At 10:52 PM 8/13/98 EDT, MICROFAB-at-aol.com wrote:
Hi All: I am looking for a sure-fire method to remove the "speckled" appearance in silicon HREM images. I've heard of etchants and other methods to remove this artifact, but I am unaware of any specific sure-fire recipes/methods. Any contributions would be greatly appreciated. Thanks in advance.
Regards,
Michael Coviello UT Arlington Materials Science Arlington, TX 817 272-5496
Does anyone have a Nikon 40X/NA 1.2 water immersion objective for sale? Any leads would be appreciated.
Most Nikon water immersion lenses for 160 mm tube models are out of stock.
Thanks, Glen
-- Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax The box said "Requires Windows95 or better". So I bought a Macintosh.
What is the source of the "speckle" to which you are referring? If it is shot noise, no etchant will have any effect. If the images have a hazy appearance due to the presence of a damaged surface layer (produced by ion milling?), you might try an alternate specimen preparation route such as chemical polishing.
++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
I suggest getting in touch with companies that make air filters (3M, for example) or your local eye/ear/nose/throat specialist. How about David Scharf?
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
I can't thank you enough for the rapid outpouring of responses to my request. With your help, I was able to generate a list of about twenty labs for my grateful customer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
Shot noise refers to poor statistical sampling in an image. For instance if N electrons arrive per pixel of the image, this will lead to a random component in the "counting" of the electrons which is of the order of root(N). Most textbooks on HREM will have a discussion of this. It is why one does not use the fastest possible film (and short exposure times) in recording HREM images, which could otherwise be done to avoid problems with drift. -Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
Speckle in HREM images? How high was the mag? Are you sure you aren't looking at the considerable ion milling radiation damage artifact that forms in Si even after only a few seconds of ion milling? I suppose the mag would have to be around 250 to 400kx, which isn't very HREM, for this radiation damage to be confused with shot noise/speckle. At lower mags the radiation damage looks like salt and pepper.
5 to 10 nm is exactly the size of the ion mill, artifactual, radiation damage, salt-and-pepper.
We try to thin specimens via tripod polishing so we don't have to ion mill for this reason. We are successful better than half the time. When we do ion mill, we do so at 2 to 3 KeV for no more than one or two minutes. We note that even 15 to 30 seconds of this low dose ion mill will create ion mill artifact. If it bothers you, don't ion mill. But, about 99% of people publishing TEMs of ion milled Si don't mention it--even those doing radiation damage studies. (OK, so I exaggerate a little).
The other responders did an excellent job of explaining shot noise, etc.
Coviello-at-mae.uta.edu on 08/14/98 03:37:15 PM Please respond to Coviello-at-mae.uta.edu To: Ronald Anderson/Fishkill/IBM-at-ibmus cc:
Hi,
Recently there has been some interest in Araldite 502 as an embedding medium. In our laboratory we have been using the mixed resin embedding which appeared in 1972 in Hayat's book "Basic Electron Microscopy Techniques". Rather than use it as a stock mixture I recalculated it to be a single mixture. (Araldite 502 30ml, Eponate 12 or LX-112 50ml, DDSA 110ml, DMP-30 1.5%, dibutyl phthalate 1/2%-2%.
Dibutyl is a plasticicer for epoxides. It does not react chemically with the monomers, but it allows for slippage along the sites of crosslinks. This influences the hardness and cuttability of the block. With a standardized 48 hour, 60deg C. polymerization time, using 1/2% dibutyl yields medium hard block, 1% gives a medium soft block, and 2% a very soft block.
The medium hard block is very good for immediate thin sectioning. The medium soft block (after 24 hrs only of polymerization at 60deg C.) is ideal for post-embedding immunogold for TEM due to its low crosslinkage. The soft block is great for endless thick sections. Glass knives last a long time, probably due to the absence of NMA. As a matter of fact, the above formulation with 1% dibutyl is the only formulation which should be employed when no diamond knives are available for thin sectioning. Araldite 502 already contains a large amount of dibutyl when purchased. That such a small variation in additions of dibutyl should make a difference in sectioning qualities has always been a mystery to me. Sometimes, an attempt is made to substitue one of the other Araldites, such as 506 for 502. The Araldites are not interchangeable. There are vast differences in composition between labels.
Any of the above mixtures can be repolymerized at 95deg C. until a block firmness of desirable qualities is achieved.
The big disadvantage of this mixture is its viscosity. Infiltration times must be lengthened over times used with epon. The mixture seperates easily and therefore must be kept well agitated. Infiltration must take place on a rotator, not on a rocker. The above formulations are unsuitable for reembedding thick sections from microscope slides. Araldite is known for its elasticity and its adhesive qualities in industry - this is counterproductive to clean seperations of sections and blocks on slides.
I have used the above formulations since 1981 for students (easy sectioning), for projects which require huge amounts of thick sections, and for immunocytochemistry-postembedding Au. They are a good tool in the laboratory. Bye, Hildy
Ken Tobin Guyot Hall Dept. of Geosciences Princeton University Princeton, NJ 08544 Ph: 609-258-1383 FAX: 609-258-1274 e-mail: tobin-at-geo.princeton.edu
I have noted with interest the several recent postings regarding the Tracor Northern ADEM SEM, its motivation and demise. Tim Moeller's postings covered the history of this development quite well, but as one who was at the center of both the development efforts and also the management decisions leading up to it, the following remarks from my perspective may be of interest.
It is generally taken as a "given" that our principal failure was that we failed to anticipate the impact on Tracor Northern EDS sales due to the reaction of the other SEM manufacturers to our entry into their market -- the so called "channel conflict". We were, in fact, deathly afraid of this scenario and I remember numerous conversations with various persons through the 70's and early 80's where I argued why it would be sheer folly for us to attempt to build our own SEM (an idea which numerous people suggested to us). So why did I change my mind, champion the effort, and ultimately give up my position as head of R&D at TN in order to lead the ADEM development team?
The decision to proceed with ADEM took place in 1983, shortly after TN's launch of the very successful TN-5500 analyzer (though the major development effort didn't kick in till spring of 1985).. Though we were enjoying great market success at that time, we saw a disconcerting future on the horizon. At that time a very large fraction of our sales were based on automating other people's microscopes -- large systems involving digital imaging and SEM automation (motorized stages, column control, etc.), together with the application software to operate them. It was commonplace to sell expensive and complex systems of which perhaps 1/3 or less of TN's revenue was due to the EDS components as such. It was in the "computerized microscope" arena where we felt that we had established a technological edge in the market. However, we perceived that this was an unstable situation and that it was only going to be a matter of time before the SEM manufacturers incorporated imaging and automation capabilities into their systems. When this occurred, we reasoned, the EDS market was going to be reduced to one of selling commodity EDS components. Rather than passively await this fate, we decided to be bold and enter the SEM market ourselves, since that is where we thought the real long-term action and opportunity lay. Though we were very anxious about the impact on our EDS sales, we felt that we were at a "window of opportunity" where we could pull this off, but that the window was sure to close. With this in mind, we decided to make our move. The final go-ahead was made with the full understanding of 'Van' Vandenheuvel (general manager of TN), Bill Buffo (president of TN and Tracor VP of the Instruments Group) and Frank McBee (president of Tracor) that this was going to have a negative impact on EDS sales, but it was felt that we could weather this as Tracor was a very healthy company at that point. Since TN had been a consistently profitable company for the past 20 years, it was Tracor's position that it was appropriate to invest short-term profits for a future position in a market where we thought we had something unique to offer.
Central to our strategy was a conviction that we could provide a substantially better value by designing a fully integrated SEM and thus offer both better price and productivity than was possible by the "piecemeal" approach of installing an EDS/automation system on a stand-alone SEM manufactured by someone else. I think we actually delivered on this rather well. Despite the fact that ADEM was indeed a rather complicated system, to be fair you need to compare it with the "glued together" systems it was designed to replace. Compared to these, ADEM was a bargain, simple to run and maintain, and highly productive. I am told that the instruments in the field have proven to be reliable, and had the development and support team remained intact, I suspect the instrument could have enjoyed a long life. So what went wrong? Having thought about it a lot in the subsequent years, I note three key "surprises" which ultimately played a large role in dooming our efforts:
(1) Introduction of field-emission SEM technology; (2) The leveraged buyout of Tracor; and (3) The outbreak of "global peace".
Field emission was a technology breakthrough we didn't anticipate. We were able to keep ADEM under wraps until its market introduction in the summer of 1987, and thus stave off erosion of EDS sales until we were ready to make our move. The introduction was quite successful, and those present at the Baltimore EMSA/MAS introduction may remember the SRO crowds it drew. There was no question that ADEM made a big splash. However, it was around this time that Hitachi introduced their field-emission SEM and this quickly became the technology where people with ADEM-sized budgets thought they should spend their SEM dollars -- particularly so in the semiconductor industry, which by now had evolved into being the principal market for high-end SEMs. We had targeted the existing thermionic SEMs of the early 80's, and felt that we competed quite well with the instruments of that type -- but field emission was a "paradigm shift" for which we were unprepared. However, we were playing for the "long term" and our development team knew we would have to work both hard and smart to pull off what we were attempting. We started work on our own field emission instrument.
About this time, the financial bottom was also dropping out of Tracor. With incredible ill-timing, a private investment group had finalized a leveraged buyout of Tracor just days before the major market crash of the late '80s. Then, the collapse of the Soviet Union depressed the defense industry on which Tracor relied for much of their revenue. Also around this time, McBee, Buffo, and Van all resigned, and with them went the commitment to the product. Instead of struggling through a "trough" as our SEM sales replaced lost EDS sales, as we expected, we found the whole company fighting for survival. Eventually, Tracor was forced to sell off the instruments group and the new management decided that they had to jettison ADEM in late 1990 -- that's when I joined Rich Lee to participate in his vision of a low-cost automatable SEM.
Could ADEM have succeeded had these "surprises" not taken place? I like to think so, but I also now see the huge obstacles we faced with a clarity that only hindsight can provide. I think there was a certain amount of "hubris" involved in our failure, and I accept the responsibility for that. There were also some engineering decisions I/we made which are questionable in retrospect. ADEM was loaded with technological innovations, but I do wish in retrospect that I had been more selective and focused -- had we kept the product simpler (and less expensive) I think we might have vastly improved our chances. (That's a hard-learned lesson that I am happy to say that we have been able to apply to the development of the PERSONAL SEM at RJ Lee Instruments -- and with a lot more satisfactory business outcome, I might add). And, of course, had we had access to the kind of powerful/inexpensive PC technology which is available today, we could have saved a whole lot of engineering and provided a lot more bang for the buck -- another lesson learned.
The whole ADEM experience did leave me with deeply conflicted feelings. On the one hand, I remain very proud of what our ADEM team accomplished, and that we had the nerve to take on what we knew was a huge challenge. But at the same time I am also painfully aware of the personal and economic costs and the "bottom line" fact that we ultimately failed to achieve our vision. The one thing I can say with certainty is that the ADEM team was spectacularly competent and, based on their dedication and effort, they deserved an outcome far better than what I was able to deliver.
A final bit of irony -- while we were laboring in our secret quarters in an industrial park miles from the main TN plant, the ADEM team occasionally noted the weird activities of a nearby startup. It turned out they were marketing collector dolls. Fast-forward seven years. I have an article on my desk from our local Pittsburgh paper describing the phenomenon that the Pleasant Company "American Girl" dolls have become -- stating that the company turned a PROFIT of $80 million last year! Mattel recently purchased the company for something north of $300 million, as I recall. ADEM has been a defunct product for years. Is there a lesson here?
Fred Schamber RJ Lee Instruments Limited ....................... mailto:fhscham-at-SGI.NET
After ~ 2 weeks of testing I am going to test the full filtering of Email to the ListServer this weekend. Any mail which triggers a Filter Flag is tagged as potential JUNK/SPAM mail and WILL NOT be posted to the ListServer. The sender of the suspect Email is sent a warning message. If you receive one of these warning messages please follow the instructions you receive so that we can sort out any remaining issues.
Remember the filter only operates on the text in the Email Header and Subject lines. It does not search the body of the text . Suspect or Bogus Email Addresses, Keywords in the Subject line (or lack of a Subject) are the some of criteria I am using.
} From the subscriber base the only problem that I expect will be the occasional rejection due to suspect Email address, this will be true if your local Email provider changes and/or modifies headers in your message which makes your mail appear to be posted from a different address. When this happens just follow the instructions and send me a confirming message, I can then add your particuliar Email address to a list of "exceptions" and your postings will go through by-passing the filter.
There are obviously no guarentee's that all junk mail will be filtered, but this should handle the majority (I hope).
If things go well, and there are no major problems over the weekend (when the mail is "light"), then I will obviously leave the filter on-line. Please report problems to me at:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have noted with interest the several recent postings regarding the } Tracor Northern ADEM SEM, its motivation and demise. Tim Moeller's } postings covered the history of this development quite well, but as one } who was at the center of both the development efforts and also the } management decisions leading up to it, the following remarks from my } perspective may be of interest. } } It is generally taken as a "given" that our principal failure was that } we failed to anticipate the impact on Tracor Northern EDS sales due to } the reaction of the other SEM manufacturers to our entry into their } market -- the so called "channel conflict". We were, in fact, deathly } afraid of this scenario and I remember numerous conversations with } various persons through the 70's and early 80's where I argued why it } would be sheer folly for us to attempt to build our own SEM (an idea } which numerous people suggested to us). So why did I change my mind, } champion the effort, and ultimately give up my position as head of R&D } at TN in order to lead the ADEM development team? } } The decision to proceed with ADEM took place in 1983, shortly after TN's } launch of the very successful TN-5500 analyzer (though the major } development effort didn't kick in till spring of 1985).. Though we were } enjoying great market success at that time, we saw a disconcerting } future on the horizon. At that time a very large fraction of our sales } were based on automating other people's microscopes -- large systems } involving digital imaging and SEM automation (motorized stages, column } control, etc.), together with the application software to operate them. } It was commonplace to sell expensive and complex systems of which } perhaps 1/3 or less of TN's revenue was due to the EDS components as } such. It was in the "computerized microscope" arena where we felt that } we had established a technological edge in the market. However, we } perceived that this was an unstable situation and that it was only going } to be a matter of time before the SEM manufacturers incorporated imaging } and automation capabilities into their systems. When this occurred, we } reasoned, the EDS market was going to be reduced to one of selling } commodity EDS components. Rather than passively await this fate, we } decided to be bold and enter the SEM market ourselves, since that is } where we thought the real long-term action and opportunity lay. Though } we were very anxious about the impact on our EDS sales, we felt that we } were at a "window of opportunity" where we could pull this off, but that } the window was sure to close. With this in mind, we decided to make } our move. The final go-ahead was made with the full understanding of } 'Van' Vandenheuvel (general manager of TN), Bill Buffo (president of TN } and Tracor VP of the Instruments Group) and Frank McBee (president of } Tracor) that this was going to have a negative impact on EDS sales, but } it was felt that we could weather this as Tracor was a very healthy } company at that point. Since TN had been a consistently profitable } company for the past 20 years, it was Tracor's position that it was } appropriate to invest short-term profits for a future position in a } market where we thought we had something unique to offer. } } Central to our strategy was a conviction that we could provide a } substantially better value by designing a fully integrated SEM and thus } offer both better price and productivity than was possible by the } "piecemeal" approach of installing an EDS/automation system on a } stand-alone SEM manufactured by someone else. I think we actually } delivered on this rather well. Despite the fact that ADEM was indeed a } rather complicated system, to be fair you need to compare it with the } "glued together" systems it was designed to replace. Compared to these, } ADEM was a bargain, simple to run and maintain, and highly productive. I } am told that the instruments in the field have proven to be reliable, } and had the development and support team remained intact, I suspect the } instrument could have enjoyed a long life. So what went wrong? Having } thought about it a lot in the subsequent years, I note three key } "surprises" which ultimately played a large role in dooming our efforts: } } (1) Introduction of field-emission SEM technology; } (2) The leveraged buyout of Tracor; and } (3) The outbreak of "global peace". } } Field emission was a technology breakthrough we didn't anticipate. We } were able to keep ADEM under wraps until its market introduction in the } summer of 1987, and thus stave off erosion of EDS sales until we were } ready to make our move. The introduction was quite successful, and } those present at the Baltimore EMSA/MAS introduction may remember the } SRO crowds it drew. There was no question that ADEM made a big splash. } However, it was around this time that Hitachi introduced their } field-emission SEM and this quickly became the technology where people } with ADEM-sized budgets thought they should spend their SEM dollars -- } particularly so in the semiconductor industry, which by now had evolved } into being the principal market for high-end SEMs. We had targeted the } existing thermionic SEMs of the early 80's, and felt that we competed } quite well with the instruments of that type -- but field emission was a } "paradigm shift" for which we were unprepared. However, we were playing } for the "long term" and our development team knew we would have to work } both hard and smart to pull off what we were attempting. We started } work on our own field emission instrument. } } About this time, the financial bottom was also dropping out of Tracor. } With incredible ill-timing, a private investment group had finalized a } leveraged buyout of Tracor just days before the major market crash of } the late '80s. Then, the collapse of the Soviet Union depressed the } defense industry on which Tracor relied for much of their revenue. Also } around this time, McBee, Buffo, and Van all resigned, and with them went } the commitment to the product. Instead of struggling through a "trough" } as our SEM sales replaced lost EDS sales, as we expected, we found the } whole company fighting for survival. Eventually, Tracor was forced to } sell off the instruments group and the new management decided that they } had to jettison ADEM in late 1990 -- that's when I joined Rich Lee to } participate in his vision of a low-cost automatable SEM. } } Could ADEM have succeeded had these "surprises" not taken place? I like } to think so, but I also now see the huge obstacles we faced with a } clarity that only hindsight can provide. I think there was a certain } amount of "hubris" involved in our failure, and I accept the } responsibility for that. There were also some engineering decisions I/we } made which are questionable in retrospect. ADEM was loaded with } technological innovations, but I do wish in retrospect that I had been } more selective and focused -- had we kept the product simpler (and less } expensive) I think we might have vastly improved our chances. (That's a } hard-learned lesson that I am happy to say that we have been able to } apply to the development of the PERSONAL SEM at RJ Lee Instruments -- } and with a lot more satisfactory business outcome, I might add). And, } of course, had we had access to the kind of powerful/inexpensive PC } technology which is available today, we could have saved a whole lot of } engineering and provided a lot more bang for the buck -- another lesson } learned. } } The whole ADEM experience did leave me with deeply conflicted feelings. } On the one hand, I remain very proud of what our ADEM team accomplished, } and that we had the nerve to take on what we knew was a huge challenge. } But at the same time I am also painfully aware of the personal and } economic costs and the "bottom line" fact that we ultimately failed to } achieve our vision. The one thing I can say with certainty is that the } ADEM team was spectacularly competent and, based on their dedication and } effort, they deserved an outcome far better than what I was able to } deliver. } } A final bit of irony -- while we were laboring in our secret quarters in } an industrial park miles from the main TN plant, the ADEM team } occasionally noted the weird activities of a nearby startup. It turned } out they were marketing collector dolls. Fast-forward seven years. I } have an article on my desk from our local Pittsburgh paper describing } the phenomenon that the Pleasant Company "American Girl" dolls have } become -- stating that the company turned a PROFIT of $80 million last } year! Mattel recently purchased the company for something north of $300 } million, as I recall. ADEM has been a defunct product for years. Is } there a lesson here? } } Fred Schamber } RJ Lee Instruments Limited } ....................... } mailto:fhscham-at-SGI.NET
Leitz, Model SM-LUX, binocular head with 10x Periplan eyepieces. The 170mm tube length objectives are: 3.5/0.10, 10/0.25, 45/0.65, and 100/1.30 oil, the 45x and 100x are spring loaded noses. All the objectives lens are Leitz. The microscope has a light source with transformer. Everything works smoothly and the optics are crisp and clear. There is a small wear spot on the frame where the paint has been rubbed away by the support inside the case. Otherwise it is in very good condition.
The sequence of ADEM postings tell a fascinating story that deserves more than the obscurity of the listserver archives. Will you consider publishing? [Are you reading this, Don Grimes?]
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
This is not a SM-LUX but a SM microscope and worth to my knowledge not more than $300.
Joseph Passero wrote:
} Leitz, Model SM-LUX, binocular head with 10x Periplan eyepieces. The } 170mm tube length objectives are: 3.5/0.10, 10/0.25, 45/0.65, and } 100/1.30 oil, the 45x and 100x are spring loaded noses. All the } objectives lens are Leitz. The microscope has a light source with } transformer. Everything works smoothly and the optics are crisp and } clear. There is a small wear spot on the frame where the paint has been } rubbed away by the support inside the case. Otherwise it is in very good } condition. } } See attached photo file. } } Entertaining Offers } } Thank You } } Joseph Passero } jp-at-spacelab.net } } ------------------------------------------------------------------------ } [Image]
Does anyone have comments on the suitability of this printer for TEM working prints? The input would be from a 1024x1024 digital camera and a SprintScan 45 from large-format negatives. A comparison with the Epson Stylus would be useful. Thanks Sally
---------------------------------------------------------------------- Dr Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: GPO Box 475 ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 (0)2 6249 2743 |Australian National Univ. FAX 61 (0)2 6249 4891 |Canberra, Australia 2601 http://online.anu.edu.au/EMU/home.htm
As a member of the ADEM development/manufacturing/service team, I would like to respond to the recent postings on this subject. As a member of Fred Schamber's team at Tracor, I would just like to say that I am grateful and honored to have had the opportunity to work with such an intense, innovative, and fun group of people. I joined the ADEM project just months before its introduction and worked as a final assembly and test technician. The knowledge I gained from that experience is inmeasurable.
In my opinion, there is no way the demise of the ADEM can be attributed to any decision or action that Fred made. There were so many external influences at play at the time that I think we never really had a chance to meet the "bottom line" in the time frame the corporation wanted. I think I speak for all the members of the ADEM team when, if asked if I'd do it all again I can only say this: In a heartbeat!
We keep our DMP-30 tightly sealed in the refrigerator and bring it to room temperature (20 deg C) in the fume hood for the 3 days of resin infiltration and embedding. Always keep lid closed when not in use, use a clean disposable glass pipette each time. Once finished store back in the refrigerator. Have stored DMP-30 for up to 2 years - discard if it appears too yellow or becomes slightly granular.
Haven't tried BDMA as have had no problems with DMP-30
Belinda White Senior Technician Centre for Electron Microscopy University of Natal Scottsville,Pietermaritzburg South Africa
If the contrast is from ion milling as Ron Anderson suggests, you might want to try the small angle cleavage technique which will eliminate this as well as the ion mixing/amorphization layers on these samples. In fact, one of the disadvantages of the technique is a lack of amorphous areas for focussing. Silicon feathers down to almost nothing and will provide you with extremely thin areas. You can produce XTEM and plan view samples using this technique. If you need site-specific preparation, this technique will not work for you. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Mike Coviello To: microscopy-at-sparc5.microscopy.com
Hi All: I am looking for a sure-fire method to remove the "speckled" appearance in silicon HREM images. I've heard of etchants and other methods to remove this artifact, but I am unaware of any specific sure-fire recipes/methods. Any contributions would be greatly appreciated. Thanks in advance.
Regards,
Michael Coviello UT Arlington Materials Science Arlington, TX 817 272-5496
Belinda White wrote: ================================================ We keep our DMP-30 tightly sealed in the refrigerator and bring it to room temperature (20 deg C) in the fume hood for the 3 days of resin infiltration and embedding. Always keep lid closed when not in use, use a clean disposable glass pipette each time. Once finished store back in the refrigerator. Have stored DMP-30 for up to 2 years - discard if it appears too yellow or becomes slightly granular.
Haven't tried BDMA as have had no problems with DMP-30 =============================================== One reason why DMP-30 has retained its popularity in many markets of the world is that it is not a HAZMAT from the standpoint of shipping. The inclusion of one bottle of BDMA, which is a HAZMAT (UN#2619), into an embedding resin kit, which otherwise does not contain any other HAZMATs, increases greatly the shipping costs, since it can not go by cheaper methods and can go only by air freight (or sea freight). Yet as was pointed out, many people get laboratory results that are just fine with DMP-30.
On the other hand, and it is not widely known, by taking advantage of certain small quantity shipping exemptions, one can still get their BDMA in such markets relatively inexpensively, but by using a different packaging approach, and this approach is explained on our website under "Hazardous Materials: Good News and Bad News". For the US domestic market, these differences are very small and are not significant. The bigger differences come into play when making shipments over international boundaries.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
As most regular subscribers know, Microscopic Explorations, a hands on science program designed to teach microscopy has recently been publishe= d by the Lawrence Hall of Science. The book is a direct result of Project M= icro and the monumental efforts of Caroline Schooley. It has many interesti= ng microscopic activities for 9-12 year old students including one activit= y where students examine sand with their microscopes. The Midwest Micros= copy and Microanalysis is working with the local science teachers to put tog= ether several kits to support this program, and we need sand from around the world. If you would like to donate a sample please dry out a handful o= f sand, seal it in a plastic bag (Ziplock will work), and mail it to:
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd Abbott Park, IL 60064-3537
Please include the location the sand was collected from. Your donation= s will be greatly appreciated. If we get enough donations we will be happy to= share them with fellow scientists who are supporting science education.
Please accept my apologies for posting this, as it is not strictly a microscopy-related question, but I was hoping that some of our esteemed subscribers would be able to offer some advice, or point me in the right direction.
We are looking at proliferating cells in animal tissues and, at present, are giving the animals (goats) 100mg/Kg body weight IV two hours prior to sacrifice. One of our researchers has suggested giving the animals another dose approx 5-7 days prior to this one.
Before we consider this, we wish to know a couple of things about BrdU, and have been unable to obtain any information from our supplier (Sigma). How long does BrdU remain in the bloodstream before being excreted, and which organ(s) is responsible for BrdU breakdown and excretion. What is the half-life of BrdU, ie, how long can the BrdU be detected immunocytochemically in cells after initial dosage? What accumulative effect will the BrdU have at this dose if given several times over a short time frame, say three weeks?
} Has anyone out there tried antigen retrieval method on EM samples?
} Qing Yang, Ph.D.
} National Institutes of Health
----------------------------------------
Dear Qing: Two References from my Microwave Methods database that may be of help!
1. Utsunomiya H, Shan L, Kawano I, Iwasaki A, Ono K, Kobayashi A, Kuma K, Kishikawa S, Kakudo K: Immunolocalization of parathyroid hormone in human parathyroid glands with special references to microwave antigen retrieval. Endocr Pathol 1995, 6:223-227
The subcellular localization of parathyroid hormone (PTH) in the normal human parathyroid glands with particular reference to microwave antigen retrieval was investigated using peroxidase-labeled PTH antibody, immunohistochemical, and immunoelectron microscopic methods. The results revealed that PTH granules existed mainly as pro-PTH on the trans side of Golgi and in the regions adjacent to Golgi apparatus. Only a small proportion of secretory granules were stored near the plasma membrane. Microwave irradiation was essential for the immunodetection of PTH. As the irradiative time extended from 1 to 30 min, the staining intensity increased, and the subcellular preservation decreased. Microwave irradiation for 15 min (with the sections in citrate buffer) with a power output of 500 W is the most ideal for PTH antigen retrieval, as well as for subcellular preservation.
2. Stirling JW, Graff PS: Antigen unmasking for immunoelectron microscopy: labeling is improved by treating with sodium ethoxide or sodium metaperiodate, then heating on retrieval medium. J Histochem Cytochem 1995, 43:115-123
To optimize the ultrastructural localization of immunoglobulin G in corneal crystalloid deposits, we compared a range of antigen unmasking techniques. A human corneal biopsy specimen was fixed in formalin, post-osmicated, and embedded in epoxy resin for electron microscopy. Thin sections were immunogold-labeled for IgG after treatment with sodium ethoxide or sodium metaperiodate. Sections were also treated by heating them at 95 degrees C while they floated on water, 0.01 M citrate buffer (pH 6.0), or sodium metaperiodate. The treatments were applied separately and combined. After labeling, crystalloids in untreated sections had a probe density of 5 particles/microns2. Crystalloids in sections treated only with sodium ethoxide or sodium metaperiodate had probe densities of 15-20 particles/microns2. Sodium ethoxide combined with heating on water, or citrate buffer, gave probe densities of 140-160 particles/microns2. Sodium metaperiodate combined with heating on citrate buffer gave the highest probe density (195 particles/microns2). Although sodium ethoxide coupled with heating increased probe density, the ethoxide etched the sections and caused unacceptable damage. Treatment with sodium metaperiodate followed by heating on citrate buffer is recommended for antigen unmasking. This combination gave a high probe density and sections remained intact, with good ultrastructural detail.
FALL 1998 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-V2)
NASSAU COMMUNITY COLLEGE, Long Island, NY
A fourteen week, fall 1998 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered ONE EVENING PER WEEK, Thursdays, starting at 5:30pm. Classes will begin on Sept. 3 and end on Dec. 10, 1998.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 per credit.
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
For those without www access, the catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students. Seats are still available for this section. The college is currently processing late registrations.
Questions regarding the actual registration process can be directed to our registrar at (516) 572-7355. ________________________________________________________________________________
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy -- 4 credits Prerequisites: BIO 109-110 or equivalent, CHE 151-152 or equivalent. An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies. ________________________________________________________________________________
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Are you interested in Digital microscopy? If so, exciting things are = happening at Sound Vision Inc. The founders of the acclaimed Leaf = Systems, the MicroLumina and other digital products have developed the = SVMicro, a new high-resolution digital camera for the microscope for = about $2000.=20
a.. The camera mounts directly to the microscope via a c-mount and = is tethered to your computer.=20 b.. The software allows instant archiving of images in a simple = click and save fashion.=20 c.. The SVMicro utilizes three shot RGB technology that gives you = selectable resolutions for various output file sizes up to 9MB. d.. It is perfect for sending images across the Internet, = documentation, archiving or high-resolution publication.=20 e.. The camera is available with ECP parallel port PC or a SCSI = connection for MAC users. f.. The same camera also takes great black and white pictures in a = one shot mode. g.. Able to do long exposures for most types of fluorescence without = any high cost cooling system h.. Able to rotate the green filter in front of the sensor to take = one shot images=20 i.. Can save sequential shots to your hard drive without leaving the = plugin. To find out more about the SVMicro Digital Camera for the microscope, = please visit our web site at www.soundvisioninc.com. You can also call = Sound Vision=92s sales department at 508-270-0044. Or send me an email = at gwray-at-soundvisioninc.com.=20
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.2106.6"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {B} {FONT size=3D2} {P} Are you interested in Digital microscopy? {/B} If so, exciting things = are=20 happening at {B} Sound Vision Inc {/B} . The founders of the acclaimed Leaf =
Systems, the MicroLumina and other digital products have developed the=20 {B} SVMicro {/B} , a new high-resolution digital camera for the microscope = for=20 about {B} $2000 {/B} . {/P} {UL} {LI} The camera mounts directly to the microscope via a c-mount and = is=20 tethered to your computer. {/LI} {LI} The software allows instant archiving of images in a simple = click and=20 save fashion. {/LI} {LI} The SVMicro utilizes three shot RGB technology that gives you = selectable=20 resolutions for various output file sizes up to 9MB. {/LI} {LI} It is perfect for sending images across the Internet, = documentation,=20 archiving or high-resolution publication. {/LI} {LI} The camera is available with ECP parallel port PC or a SCSI = connection=20 for MAC users. {/LI} {LI} The same camera also takes great black and white pictures in a = one shot=20 mode. {/LI} {LI} Able to do long exposures for most types of fluorescence without = any=20 high cost cooling system {/LI} {LI} Able to rotate the green filter in front of the sensor to take = one shot=20 images {/LI} {LI} Can save sequential shots to your hard drive without leaving the =
plugin. {/LI} {/UL} {P} To find out more about the SVMicro Digital Camera for the microscope, = please=20 visit our web site at {/FONT} {A = href=3D"http://www.soundvisioninc.com/"} {FONT=20 size=3D2} www.soundvisioninc.com {/FONT} {/A} {FONT size=3D2} . You can also = call Sound=20 Vision’s sales department at 508-270-0044. Or send me an email at=20 {/FONT} {A href=3D"mailto:gwray-at-soundvisioninc.com"} {FONT=20 size=3D2} gwray-at-soundvisioninc.com {/FONT} {/A} {U} {FONT size=3D2} . {/P} {/U} {P} {/P} {P} Best Regards, {/P} {P} {/P} {P} Gary Ray {/P} {P} Sales Department {/P} {P} Sound Vision Inc. {/P} {/FONT} {/DIV} {/BODY} {/HTML}
This is a multi-part message in MIME format. --------------C13D3920A222C3F36012EB4A Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Just a personal note of thanks for filling in the gaps, Fred! I agree completely with Caroline Schooley's statement that these posts tell a fascinating story that deserve more than the obscurity of the listserver archives. I would take issue with only one thing you said -- indeed, you take too much upon yourself by saying that the ADEM team "deserved an outcome far better than what I was able to deliver." Of course, anybody who knows your modest and generous nature would almost expect you to say such a thing, accepting responsibility where no real culpability exists, but everyone here should know that you did us no wrong. Certainly it was regrettable, but no one blames you personally for the outcome brought about by factors quite beyond your (or anyone else's) control. To the contrary, I'm sure many team members feel as I do that it was a tremendous privlege to have been included at all, and look back with fond memories on that once-in-a-lifetime experience. We took our best shot, and lost -- but we almost won, and would have won big. That was the chance we took; sometimes you get the bear and sometimes the bear gets you. Besides, there were other personal benefits accrued through the experience, despite the outcome. Anyway, thanks again!
Fred Schamber wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have noted with interest the several recent postings regarding the } Tracor Northern ADEM SEM, its motivation and demise. Tim Moeller's } postings covered the history of this development quite well, but as one } who was at the center of both the development efforts and also the } management decisions leading up to it, the following remarks from my } perspective may be of interest. } } It is generally taken as a "given" that our principal failure was that } we failed to anticipate the impact on Tracor Northern EDS sales due to } the reaction of the other SEM manufacturers to our entry into their } market -- the so called "channel conflict". We were, in fact, deathly } afraid of this scenario and I remember numerous conversations with } various persons through the 70's and early 80's where I argued why it } would be sheer folly for us to attempt to build our own SEM (an idea } which numerous people suggested to us). So why did I change my mind, } champion the effort, and ultimately give up my position as head of R&D } at TN in order to lead the ADEM development team? } } The decision to proceed with ADEM took place in 1983, shortly after TN's } launch of the very successful TN-5500 analyzer (though the major } development effort didn't kick in till spring of 1985).. Though we were } enjoying great market success at that time, we saw a disconcerting } future on the horizon. At that time a very large fraction of our sales } were based on automating other people's microscopes -- large systems } involving digital imaging and SEM automation (motorized stages, column } control, etc.), together with the application software to operate them. } It was commonplace to sell expensive and complex systems of which } perhaps 1/3 or less of TN's revenue was due to the EDS components as } such. It was in the "computerized microscope" arena where we felt that } we had established a technological edge in the market. However, we } perceived that this was an unstable situation and that it was only going } to be a matter of time before the SEM manufacturers incorporated imaging } and automation capabilities into their systems. When this occurred, we } reasoned, the EDS market was going to be reduced to one of selling } commodity EDS components. Rather than passively await this fate, we } decided to be bold and enter the SEM market ourselves, since that is } where we thought the real long-term action and opportunity lay. Though } we were very anxious about the impact on our EDS sales, we felt that we } were at a "window of opportunity" where we could pull this off, but that } the window was sure to close. With this in mind, we decided to make } our move. The final go-ahead was made with the full understanding of } 'Van' Vandenheuvel (general manager of TN), Bill Buffo (president of TN } and Tracor VP of the Instruments Group) and Frank McBee (president of } Tracor) that this was going to have a negative impact on EDS sales, but } it was felt that we could weather this as Tracor was a very healthy } company at that point. Since TN had been a consistently profitable } company for the past 20 years, it was Tracor's position that it was } appropriate to invest short-term profits for a future position in a } market where we thought we had something unique to offer. } } Central to our strategy was a conviction that we could provide a } substantially better value by designing a fully integrated SEM and thus } offer both better price and productivity than was possible by the } "piecemeal" approach of installing an EDS/automation system on a } stand-alone SEM manufactured by someone else. I think we actually } delivered on this rather well. Despite the fact that ADEM was indeed a } rather complicated system, to be fair you need to compare it with the } "glued together" systems it was designed to replace. Compared to these, } ADEM was a bargain, simple to run and maintain, and highly productive. I } am told that the instruments in the field have proven to be reliable, } and had the development and support team remained intact, I suspect the } instrument could have enjoyed a long life. So what went wrong? Having } thought about it a lot in the subsequent years, I note three key } "surprises" which ultimately played a large role in dooming our efforts: } } (1) Introduction of field-emission SEM technology; } (2) The leveraged buyout of Tracor; and } (3) The outbreak of "global peace". } } Field emission was a technology breakthrough we didn't anticipate. We } were able to keep ADEM under wraps until its market introduction in the } summer of 1987, and thus stave off erosion of EDS sales until we were } ready to make our move. The introduction was quite successful, and } those present at the Baltimore EMSA/MAS introduction may remember the } SRO crowds it drew. There was no question that ADEM made a big splash. } However, it was around this time that Hitachi introduced their } field-emission SEM and this quickly became the technology where people } with ADEM-sized budgets thought they should spend their SEM dollars -- } particularly so in the semiconductor industry, which by now had evolved } into being the principal market for high-end SEMs. We had targeted the } existing thermionic SEMs of the early 80's, and felt that we competed } quite well with the instruments of that type -- but field emission was a } "paradigm shift" for which we were unprepared. However, we were playing } for the "long term" and our development team knew we would have to work } both hard and smart to pull off what we were attempting. We started } work on our own field emission instrument. } } About this time, the financial bottom was also dropping out of Tracor. } With incredible ill-timing, a private investment group had finalized a } leveraged buyout of Tracor just days before the major market crash of } the late '80s. Then, the collapse of the Soviet Union depressed the } defense industry on which Tracor relied for much of their revenue. Also } around this time, McBee, Buffo, and Van all resigned, and with them went } the commitment to the product. Instead of struggling through a "trough" } as our SEM sales replaced lost EDS sales, as we expected, we found the } whole company fighting for survival. Eventually, Tracor was forced to } sell off the instruments group and the new management decided that they } had to jettison ADEM in late 1990 -- that's when I joined Rich Lee to } participate in his vision of a low-cost automatable SEM. } } Could ADEM have succeeded had these "surprises" not taken place? I like } to think so, but I also now see the huge obstacles we faced with a } clarity that only hindsight can provide. I think there was a certain } amount of "hubris" involved in our failure, and I accept the } responsibility for that. There were also some engineering decisions I/we } made which are questionable in retrospect. ADEM was loaded with } technological innovations, but I do wish in retrospect that I had been } more selective and focused -- had we kept the product simpler (and less } expensive) I think we might have vastly improved our chances. (That's a } hard-learned lesson that I am happy to say that we have been able to } apply to the development of the PERSONAL SEM at RJ Lee Instruments -- } and with a lot more satisfactory business outcome, I might add). And, } of course, had we had access to the kind of powerful/inexpensive PC } technology which is available today, we could have saved a whole lot of } engineering and provided a lot more bang for the buck -- another lesson } learned. } } The whole ADEM experience did leave me with deeply conflicted feelings. } On the one hand, I remain very proud of what our ADEM team accomplished, } and that we had the nerve to take on what we knew was a huge challenge. } But at the same time I am also painfully aware of the personal and } economic costs and the "bottom line" fact that we ultimately failed to } achieve our vision. The one thing I can say with certainty is that the } ADEM team was spectacularly competent and, based on their dedication and } effort, they deserved an outcome far better than what I was able to } deliver. } } A final bit of irony -- while we were laboring in our secret quarters in } an industrial park miles from the main TN plant, the ADEM team } occasionally noted the weird activities of a nearby startup. It turned } out they were marketing collector dolls. Fast-forward seven years. I } have an article on my desk from our local Pittsburgh paper describing } the phenomenon that the Pleasant Company "American Girl" dolls have } become -- stating that the company turned a PROFIT of $80 million last } year! Mattel recently purchased the company for something north of $300 } million, as I recall. ADEM has been a defunct product for years. Is } there a lesson here? } } Fred Schamber } RJ Lee Instruments Limited } ....................... } mailto:fhscham-at-SGI.NET --------------C13D3920A222C3F36012EB4A Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Timothy Moeller Content-Disposition: attachment; filename="vcard.vcf"
Hello to those in listserv land - I'm a long time lurker (two or more years) first time poster. There have been bunches of good stuff on the list that have saved me time, money and effort, for which, thanks to all. Now, I actually have some questions that I have not seen addressed as yet. A bit of background on me: I manage Arizona State University's Life Science EM Lab and have for over twenty years. We are a teaching and research lab with a STEM/EDS, a TEM, and an old, non-functional SEM which is the crux of all my questions. We are writing a grant proposal for a replacement for the SEM and have been very interested in a Philips/FEI ESEM FEG for its apparent resolution, flexibility, and all round usefulness. The questions we have are: 1) Can work be done on unfixed, hydrated biological specimens without cell walls such as animal tissue and/or cultured cells? 2) Can this type of work be done at high resolution, either at high or low KeV and how much beam damage does one expect? 3) Is manipulation of water vapor pressure / temperature (w/Peltier stage) sufficiently precise to do this type of work routinely? 4) Do unprotected cells simply explode when introduced into the scope? 5) Does anyone out there have such a scope in a biologically based lab or know someone who does?? 6) We have searched the literature and found virtually NO bio based papers in our meanderings. Are there papers we are missing? (we do know that FEG ESEM is new and not many are about - how about the older Electroscan W filament or LaB6 types? Are they amenable to high-ish resolution work on bio stuff? OK. TOO much bandwidth for a first post. Thanks in advance for any and all info or leads. --------------------------------------------------------------- William P. Sharp Department of Botany Arizona State University Tempe, AZ 85287-1601 --------------------------------------------------------------- voice (602) 965-3210 | fax (602)965-6899 | email wsharp-at-asu.edu
Sorry, Mr.Passero, This community doesn't need to see an attached photo of a microscope. Please use your computer for better purposes.........
Joseph Passero wrote:
} Leitz, Model SM-LUX, binocular head with 10x Periplan eyepieces. The } 170mm tube length objectives are: 3.5/0.10, 10/0.25, 45/0.65, and } 100/1.30 oil, the 45x and 100x are spring loaded noses. All the } objectives lens are Leitz. The microscope has a light source with } transformer. Everything works smoothly and the optics are crisp and } clear. There is a small wear spot on the frame where the paint has been } rubbed away by the support inside the case. Otherwise it is in very good } condition. } } See attached photo file. } } Entertaining Offers } } Thank You } } Joseph Passero } jp-at-spacelab.net } } ------------------------------------------------------------------------ } [Image]
This posting is only relevant to those who from time-to-time import chemicals from overseas. Chuck Garber's posting is rather similar to one he produced here about a year ago. Dangerous Goods shipping rules are no secret to any EM supplier. The "shipping DG's in limited quantity" rules are of little use, in fact BDMA is about the only chemical were one could save the DG fee, but it still must be airfreighted. Note A$1 = US$0.58
Here is a part of our dangerous goods notes from our online - Dangerous goods in limited quantities: Most chemicals can be shipped overseas in limited quantities. This rule allows up to 500ml or grams but packed in 30ml/g lots. Only two of our DG chemicals, osmium tetroxide and propylene oxide, cannot be shipped under that clause; for shipping, these two chemicals are always treated as dangerous goods. All our other DG chemicals could be airfreighted to overseas destinations, but never posted. However, it is not viable to ship 500g quantities dispensed in 30g quantities. Hence this rule is only useful for a few items, for instance BDMA, which is normally shipped in 100ml quantities. This chemical is also available as 4x25ml for overseas customers. When shipping DGs in limited quantities, the saving is only the dangerous goods fee, which is currently A$60. We would still need to charge for airfreight (whereas we Post (orders } $50) for free.
Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes **************************** www.proscitech.com.au *****
On Tuesday, 18 August 1998 0:17, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: ..... ...... On the other hand, and it is not widely known, by taking } advantage of } certain small quantity shipping exemptions, one can still } get their BDMA in } such markets relatively inexpensively, but by using a } different packaging } approach, and this approach is explained on our website } under "Hazardous } Materials: Good News and Bad News". For the US domestic } market, these } differences are very small and are not significant. The } bigger differences } come into play when making shipments over international } boundaries. } } Chuck
William Chris Gilpin {cgilpin-at-fs1.sem.man.ac.uk} is probably the most experienced biological ESEM user and I know he sometimes posts to this list so I expect you will hear from him. I hope he provides us all with a sample of references to bio ESEM. I suggest you contact Trisha Rice at Electroscan {trice-at-electroscan.com} as she is probably the most experienced at ESEM FEG and will be able to answer your FEG specific questions. Unfixed hydrated biologicals can be examined with enough control to keep the specimen hydrated. We have a LaB6 ESEM and do mainly materials and particles with resolution comparable to conventional SEM, I suspect the same is true for biologicals. Good luck, Scott
Not an endorsement, no financial stake, just a satisfied customer.
} interested in a Philips/FEI ESEM FEG for its apparent resolution, } flexibility, and all round usefulness. The questions we have are: } 1) Can work be done on unfixed, hydrated biological specimens without cell } walls such as animal tissue and/or cultured cells? } 2) Can this type of work be done at high resolution, either at high or low } KeV and how much beam damage does one expect? } 3) Is manipulation of water vapor pressure / temperature (w/Peltier stage) } sufficiently precise to do this type of work routinely? } 4) Do unprotected cells simply explode when introduced into the scope? } 5) Does anyone out there have such a scope in a biologically based lab or } know someone who does?? } 6) We have searched the literature and found virtually NO bio based papers } in our meanderings. Are there papers we are missing? (we do know that FEG } ESEM is new and not many are about - how about the older Electroscan W } filament or LaB6 types? Are they amenable to high-ish resolution work on } bio stuff?
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
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A Postdoctoral position in electron microscopy is available at Unilever Research Port Sunlight Laboratory Wirral UK.
Qualifications: Recent PhD in Material Science or related area. High level of hands-on experience in TEM and proficient ultramicrotomy expertise are essential for success in this role. Cryo experience at this level is desirable. Excellent communication, interpersonal and team working skills required.
The position is for one year in the first instance.
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I too would appreciate information on this subject. Please post responses to the listserve or, Bill, would you mind summarizing responses and then posting the summary?
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
Hello to those in listserv land - I'm a long time lurker (two or more years) first time poster. There have been bunches of good stuff on the list that have saved me time, money and effort, for which, thanks to all. Now, I actually have some questions that I have not seen addressed as yet. A bit of background on me: I manage Arizona State University's Life Science EM Lab and have for over twenty years. We are a teaching and research lab with a STEM/EDS, a TEM, and an old, non-functional SEM which is the crux of all my questions. We are writing a grant proposal for a replacement for the SEM and have been very interested in a Philips/FEI ESEM FEG for its apparent resolution, flexibility, and all round usefulness. The questions we have are: 1) Can work be done on unfixed, hydrated biological specimens without cell walls such as animal tissue and/or cultured cells? 2) Can this type of work be done at high resolution, either at high or low KeV and how much beam damage does one expect? 3) Is manipulation of water vapor pressure / temperature (w/Peltier stage) sufficiently precise to do this type of work routinely? 4) Do unprotected cells simply explode when introduced into the scope? 5) Does anyone out there have such a scope in a biologically based lab or know someone who does?? 6) We have searched the literature and found virtually NO bio based papers in our meanderings. Are there papers we are missing? (we do know that FEG ESEM is new and not many are about - how about the older Electroscan W filament or LaB6 types? Are they amenable to high-ish resolution work on bio stuff? OK. TOO much bandwidth for a first post. Thanks in advance for any and all info or leads. --------------------------------------------------------------- William P. Sharp Department of Botany Arizona State University Tempe, AZ 85287-1601 --------------------------------------------------------------- voice (602) 965-3210 | fax (602)965-6899 | email wsharp-at-asu.edu
RFC822 header -----------------------------------
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Dear Don,
To the contrary: a photo of a microscope often helps in identifying it and therefore in making an educated decision.
I would like to annonce and post the following new open position in the Analytical Instrumentation Facility at NCSU. Please note that this position is in addition to the position recently posted, which I have copied below for clarity. With the two microscopy positions open, we hope to find complenentary individuals to complement the expertise and skills of our existing staff. More details of our lab facilities and program can be at http://spm.aif.ncsu.edu/aif/index.html
Research Assistant / Microscopist
Announcing an opening as of Oct. 1, 1998 for the postion of Research Assistant at the North Carolina State University Analytical Instrumentation Facility (AIF).
Qualifications must include an BS as the minimum degree with higher degree desired or equivalent experience in a materials related discipline (non biological) along with hands on experience with optical metallographs, XRF, XRD, SEM, SEM/EDS, TEM and sample preparation instruments such as sample coaters, ion millers, mounting presses and grinding, polishing and sectioning instruments. Required skills include: demonstrated analytical problem solving capabilities in a multiuser analytical facility environment, extensive hands on experience with the above techniques along with their tools and accessories (e.g. metallographic, TEM, XRF, XRD and other specimen preparation, analysis of metallographic and other samples, etc.); teaching/user training; and familiarity with modern instrumentation, computer systems and experience with vacuum systems. Responsibilities include: operation and maintenance of optical metallographs, ion millers, X-Ray diffractometers and sample preparation devices such as mounting presses and grinding, polishing and sectioning devices, etc; scheduling of access to and oversight of the above instrumentation; and user training and assistance. Other responsibilities include operation of SEM and TEM and assistance with the teaching of electron microscopy laboratory classes and assistance with other graduate level engineering classes. Qualifications must include an BS as the minimum degree with higher degree desired or equivalent experience in a materials related discipline (non biological) along with hands on analytical experience in a multiuser analytical facility environment,
Please send resume and names of references to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531
North Carolina State University is an Equal Opportunity, Affirmative Action Educator and Employer. Minority and Female Applicants are especially encouraged.
Previously posted Position Open -- (still avaliable)
Microscopy and Microanalysis Research Assistant
An immediate position is open for an SEM and TEM microscopist at the North Carolina State University Analytical Instrumentation Facility (AIF) as a retirement replacement.
Duties and responsibilities include: Teaching of laboratories; user training and assistance; and instrumentation development, modification and maintenance; and analysis of a wide variety of samples. An MS or equivalent experience in a materials related discipline along with three or more years hands on experience with SEM and/or TEM is required. Required skills include: extensive hands on experience with SEM, TEM and related techniques and accessories (e.g. specimen preparation and associated tools such as evaporators, etc.); teaching/user training; and familiarity with modern electronics, computer systems and experience with vacuum systems.
Please send resume and three letters of reference to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531 or email PRUSSELL-at-NCSU.EDU.
North Carolina State University is an Equal Opportunity, Affirmative Action Educator and Employer. Minority and Female Applicants are especially encouraged.
-- Phillip E. Russell Professor, Materials Science and Engineering Director, Analytical Instrumentation Facility Box 7531, Room 318 EGRC 1010 Main Campus Drive North Carolina State University Raleigh, NC 27695-7531
Ronnie Houston wrote: } } We are looking at proliferating cells in animal tissues and, at present, are giving the animals (goats) 100mg/Kg body weight IV two hours prior to sacrifice.
This dose may be cytotoxic, it is in mice.
} One of our researchers has suggested giving the animals } another dose approx 5-7 days prior to this one.
How will that help?
} Before we consider this, we wish to know a couple of things about BrdU, } and have been unable to obtain any information from our supplier } (Sigma). How long does BrdU remain in the bloodstream before being } excreted,
BrDU is no longer available within one hour, probably unavailable within 30 minutes in mice. That can be a good thing depending on your experimental design.
} and which organ(s) is responsible for BrdU breakdown and } excretion.
My guess would be liver and/or kidney but it might be broken down by the lungs as well. Might vary with the species.
} What is the half-life of BrdU, ie, how long can the BrdU be } detected immunocytochemically in cells after initial dosage?
I don't think half-life is the correct term but cells hold on to BrDU for a long time. It will be diluted below the point of IHC detection after 2 or maybe 3 cell divisions.
} What accumulative effect will the BrdU have at this dose if given several times over a short time frame, say three weeks?
Might slow the cell cycle in targeted cells or even kill them.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I need to do some SEM imaging of uncoated/unexposed photoresist at low kV. Does anyone have any recipies or advice on this? This is on a PMMA coated Si wafer. Thanks Bill Neill
Does anyone know where I can find a positioner for microscope specimens with an overall travel range of a few microns and step size in the nanometer range?
****************************** Jim Haley e-mail: haley-at-i-cubeinc.com ******************************
William and the list, My thanks firstly to Scott for the recommend. I have been using a tungsten ESEM for biological work for the last 6 years. Although I have published little in print I have presented at numerous meetings. So only abstracts in the literature I'm afraid. OK a couple of papers on EDX in ESEM and some on pharmaceutical applications but not strictly biological. Now to your questions!
1) Yes you can work on unfixed hydrated samples. 2) depends on what you call high resolution 3) manipulation of hydration is easy with a little practice. 4) no cells do not explode. 5) Yes, I have a tungsten E3 and have used a.n.other FEG ESEM for biology. 6) See my opening my remarks
Those are the simple answers! To adequately cover the detailed answers would take up much space. Here are my main thoughts for other list readers. I can give a fuller account off list if it will help.
Cells often look very different when examined by ESEM compared to Hi VAC. I have always taken this to represent a "truer" view of biological surfaces - very smooth with little fine topography. I assume that extracellular matrix is well preserved with ESEM giving this appearance. Hence the comment on resolution. The microscope is capable of matching Hi Vac for resolution but many biological structures do not have that much detail to show. Aesthetically disappointing but with correct interpretation valuable information on structure. Low KV is difficult with a tungsten ESEM but not impossible. I routinely image at 5KV. Modification of the detector (as per Brendon Griffin)allows imaging at a few hundred volts. Beam scattering in the chamber gas is the problem and simple physics shows that the more electrons you start with (FEG } LaB6 } W)) the more will reach the sample. You are right to highlight hydration as being important. Very fine control is easy with the older instruments but I worry about the vacuum increment steps in the windows driven new instruments. 0.1 Torr or 10 Pa is not really fine enough but still possible. I have come to the conclusion that many cells are best viewed hydrated but after a short fixation in glutaraldehyde and rinse in water. This has a number of benefits. Fresh samples which arrive at 5 o'clock on a Friday can be examined on Monday morning! Beam damage is greatly reduced. Finally cells cannot be examined from culture medium or buffers because the salts precipitate out following excess liquid removal obscuring anything of interest. Unfixed samples can be examined but my experience has shown that the structure is pretty similar in both cases. It is dehydration that causes the problems not fixation. Remember that cryo stages are also available for ESEM - just use different chamber gas (see papers from the Cavendish group in Cambridge). Cells tolerate the vacuum (at least 4.6 Torr to be hydrated above freezing) well. Electroscan/FEI/Philips have video on moving insects in their promotional material.
One final point to make. If you have an ESEM you also have a very capable Hi Vac and controlled pressure instrument as well. Don't fall into the trap of should I get ESEM OR conventional - you can have both in one instrument! Another final point! The images obtained in ESEM mode should be taken as another piece of information in a bigger picture and should be seen as complementary to other SEM techniques.
My own opinions as a satisfied customer (how about a free upgrade Ralph?)
Chris Gilpin Experimental Officer Biological Sciences EM Unit University of Manchester Oxford Road Manchester M13 9PT Phone +44 (0)161 275 5170 Fax +44 (0)161 275 5171
---------------------------------------------------------------------------- ----------------- William P. Sharp wrote
We are writing a grant proposal for a replacement for the SEM and have been very interested in a Philips/FEI ESEM FEG for its apparent resolution, flexibility, and all round usefulness. The questions we have are: 1) Can work be done on unfixed, hydrated biological specimens without cell walls such as animal tissue and/or cultured cells? 2) Can this type of work be done at high resolution, either at high or low KeV and how much beam damage does one expect? 3) Is manipulation of water vapor pressure / temperature (w/Peltier stage) sufficiently precise to do this type of work routinely? 4) Do unprotected cells simply explode when introduced into the scope? 5) Does anyone out there have such a scope in a biologically based lab or know someone who does?? 6) We have searched the literature and found virtually NO bio based papers in our meanderings. Are there papers we are missing? (we do know that FEG ESEM is new and not many are about - how about the older Electroscan W filament or LaB6 types? Are they amenable to high-ish resolution work on bio stuff? OK. TOO much bandwidth for a first post. Thanks in advance for any and all info or leads. ---------------------------------------------------------------
Sodium borohydride, glycine, ammonium chloride are all used for pretreatment of sections for LM and TEM immunocytochemistry. Does anyone know the difference in actions between these? Are there any preferences for better ultrasctructural preservation among the three? I cannot seem to find this answer anywhere. Would anyone know? Or know where to find the answer?
I agree with Barbara. It would appear that one of the major on-line laboratory equipment reseller services also agrees - see www.labx.com - where most equipment advertisements are accompanied by a picture. The latter aspect is especially important at the labx site in the selling of microscopes as there are often several of the same scope, or more importantly several variants of a model for sale at the same time. However, I do agree that we do not need, and should not be subjected to unsolicited (file) attachments to e-mail.
Winton Cornell
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
Are there any Soil Scientists out there that can help my boss determine = the best way to pH the soil from his recently purchased farm. We were = trying to determine if adding tap water, dionized water, or a standard pH = buffered solution to the dry soil could influence an accurate pH. I know = this isn't rocket science, but I am sure there is a proper way to do this = and a lot of ways to get bad data. Thanks Linda Fox lfox1-at-wpo.it.luc.edu
Leica CLSM - confocal microscope (inverted). The system includes: a Leitz Fluovert inverted microscope, objective lenses, filters, krypton/argon laser, Motorola computer (running OS9) with Leica ScanWare software, and 3 monitors. Also for sale is a Focus Graphics image recorder. All reasonable offers will be considered.
If interested, please contact:
Scott Henderson, Ph.D. Director of Microscopy, Dept. Cell Biology & Anatomy, Mount Sinai School of Medicine, Box 1007, 1 Gustave L. Levy Pl., New York, NY 10029-6574
Scott Henderson, Ph.D. Director of Microscopy, Department of Cell Biology & Anatomy, Mount Sinai School of Medicine, Box 1007, One Gustave L. Levy Place, New York, NY 10029-6574
We have had pretty decent luck using an accelerating voltage around 600 - 800 eV. That is about the extent of our recipe.
John Staman Analytical Lab LSI Logic formerly Symbios Inc.
} -----Original Message----- } From: Bill Neill [SMTP:billneill-at-csi.com] } Sent: Tuesday, August 18, 1998 4:31 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Imaging of Photoresist } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need to do some SEM imaging of uncoated/unexposed photoresist at low kV. } Does anyone have any recipies or advice on this? This is on a PMMA coated } Si } wafer. } Thanks } Bill Neill } }
Dear Bill, John Cole of Hitachi Canada gave me a nice sheet of: "SEM Specimen Charging Threshold Energies of Polymers and Semiconductor Materials", which lists the upper limit for PMMA resist as 1.60 kV. This means you should not get charging of this material if you stay below this accelerating voltage. You can contact him for the complete list at: http://nsctoronto.com/ or: e-Mail nscsales-at-nsctoronto.com . You wrote: } } I need to do some SEM imaging of uncoated/unexposed photoresist at low kV. } Does anyone have any recipies or advice on this? This is on a PMMA coated Si } wafer. } Thanks } Bill Neill } Best of luck, Mary Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I suppose the addition of the photo does slow down the message time, but I was glad to see it. I did'nt get into this thread until after the first message so I do not know how long the message took to come down, but since I had an interest in seeing the unit I had the person send me a copy of it to my mailbox. As they say, A paicture is worth a thousand words and although I probably remember more models of microscopes by their given names my memory is not perfect. FOr some reason I though te sm lux was a white or grey colored newer microscope so by getting the photo it let me know what I was looking at. Perhaps in the frame of not having slow messages it is best to request the photo from the sender? If it was a small photo the transfer time wopuld not be long. Perhaps with the initial message just a small thrubnail and then request from the sender a large fit to print and frame over our desk version on request.
I am rambling so I will quit and go drink half a pot of coffee.
I will like to know the full use and range for testing materials using a = scanning electron microscope model no. JSM-35C,at present = microstructure, particle morphology and micrographs are being carried = out.We will like to know the other tests we can carry out.=20
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{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.71.2016.0"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} I will like to know the full use and = range for=20 testing materials using a scanning electron microscope model no. = JSM-35C,at=20 present microstructure, particle morphology and micrographs are being = carried=20 out.We will like to know the other tests we can carry=20 out. {/FONT} {/DIV} {/BODY} {/HTML}
} Date: Tue, 18 Aug 1998 14:00:38 -0600 (MDT) } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } To: postmessage {Microscopy-at-sparc5.microscopy.com} } Subject: Naborohydride, Glycine, A.Chl???? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi, } } Sodium borohydride, glycine, ammonium chloride are all used for } pretreatment of sections for LM and TEM immunocytochemistry. Does anyone } know the difference in actions between these? Are there any preferences } for better ultrasctructural preservation among the three? I cannot seem } to find this answer anywhere. Would anyone know? Or know where to find } the answer? } } Thank you, } Hildy } {hcrowley-at-du.edu}
Hi Hildy, et al.
NaBH4 is used as an etching agent on epoxy sections to clear away some of the resin and expose antigens for immunolabeling. (Not sure how used in LM). Trouble is, it can eat away your antigen--and even your section if too strong or left too long. Worked OK for me at ~1% for ~ 10 min, but method requires presence of lots of antigen and strong antibody. I prefer acrylic resins which don't have to be etched, or even better ultrathin cryosections--which we routinely do now.
Glycine and NH4Cl are used to quench the aldehydes before immunolabeling--supposedly frees up antigens for more access by antibodies. I don't remember the glycine conc, but the NH4Cl is usually used between 50-100 mM for about 10 min (on sections--may differ for LM??).
Cheers, S
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I will like to know the full use and range in testing materials on an = scanning electron microscope model no. JSM-35C.At present tests that can = be carried out are Microstructure, Particle Morphology and Micrographs.=20
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In addition, sodium borohydride is also a strong reducing agent, and can be used on fixed tissue to reduce aldehyde groups, thereby diminishing autofluorescence. In fact, it is a stronger reducing agent than the other two, so it can be used when they are inadequate to the task. A very good article on its use is:
Eldred WD, Zucker C, Karten HJ, Yazulla S. 1983. Comparison of fixation and penetration enhancement techniques for use in ultrastructural immunocytochemistry. J Histochem Cytochem 31:285-292.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
} On Tue, 18 Aug 1998, HILDEGARD CROWLEY wrote: } } } Date: Tue, 18 Aug 1998 14:00:38 -0600 (MDT) } } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } } To: postmessage {Microscopy-at-sparc5.microscopy.com} } } Subject: Naborohydride, Glycine, A.Chl???? } } } } Hi, } } } } Sodium borohydride, glycine, ammonium chloride are all used for } } pretreatment of sections for LM and TEM immunocytochemistry. Does anyone } } know the difference in actions between these? Are there any preferences } } for better ultrasctructural preservation among the three? I cannot seem } } to find this answer anywhere. Would anyone know? Or know where to find } } the answer? } } } } Thank you, } } Hildy } } {hcrowley-at-du.edu} } } Hi Hildy, et al. } } NaBH4 is used as an etching agent on epoxy sections to clear away some of } the resin and expose antigens for immunolabeling. (Not sure how used in } LM). Trouble is, it can eat away your antigen--and even your section if } too strong or left too long. Worked OK for me at ~1% for ~ 10 min, but } method requires presence of lots of antigen and strong antibody. I } prefer acrylic resins which don't have to be etched, or even better } ultrathin cryosections--which we routinely do now. } } Glycine and NH4Cl are used to quench the aldehydes before } immunolabeling--supposedly frees up antigens for more access by } antibodies. I don't remember the glycine conc, but the NH4Cl is usually } used between 50-100 mM for about 10 min (on sections--may differ for } LM??). } } Cheers, } S } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735
We are attempting to fix chick forebrain neurons for TEM. Several cells have been processed and embedded in Marglass. A graded series was not used to dehydrate or embed, the blocs had serious infiltration problems. To solve this I suggested using a graded series (with EtOH) and Spurtol. They tried and the Spurtol etched the plates. The plates are poly-L-lysine coated tissue culture dishes made by Falcon (Becton Dickinson). Since the Spurtol was a problem, they have been experimenting with Marglass and a graded EtOH series. However, when they try this, the plates are again etched. Any suggestions?? Change resin? Change plates? suggestions on infiltration timing would also be helpful
Thanks... Sally
Sally Burns Center for Electron Optics B5 Pesticide Research Center (517) 355-5004
I am about to do some LM autoradiographic studies and would like to know if anyone has any experience with chemography effects when sections are pre-stained with Safranin O. In the past I have successfully used Hematoxylin and PAS but prefer Hematoxylin + Saf O these days because I get a more intense staining. I am aware, however, that some dyes (e.g., Tol Blue) can't be used in pre-staining sections for autoradiography due to chemography. To save time and emulsion, I would appreciate hearing from anyone who has either successfully or unsuccessfully used Saf O prior to autoradiography. TIA Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Postdoctoral Position in Environmental Catalysis/HREM
A postdoctoral position is available at Northwestern University to work on Environmental Catalysis. The work will involve in-situ work using a unique UHV-HREM and gas treatment of various samples (see http://www.numis.nwu.edu for some information about the hardware). A strong background in HREM and crystallography is highly desirable, and experience in catalysis and/or surface science will be significant; extensive experience in other types of TEM are an alternative. To apply, send an email to L. D. Marks at ldm3-at-apollo.numis.nwu.edu including a CV and the names of referees.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 email: ldm3-at-apollo.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Ron Anderson has asked that I request a copy of of the complete list (of upper limits for PMMA resist). Please mail it to the following:
Ronald M. Anderson Treasurer, Microscopy Society of America c/o I.B.M. Corporation Analytical Services Group 1580 State Route 52, Mail Stop E40 Hopewell Jct., NY 12533-6531
Thank you so much .
Regards,
Colleen Marie Genadry Mail Stop E40, B/630, Dept. 13WA Lotus Notes ID: IBMUSM08(CGENADRY) VM ID: FSHVM1(CGENADRY) T/L532-2664 } From Outside call (914)-892-2664 FAX #: (914)-892-2003
I'm looking for a company that recoats phosphorus TEM screens. In particular, a company that uses a greener phosphorus than more yellow. In the past I used a company called Grant Scientific but have been unable to locate them.
Does anyone know how to get commercially available actin to disperse as F-actin? We have not had good luck getting a lyophilyzed powder of porcine actin (Sigma) into solution as dispersed F-actin filaments. We usually get quite a bit of clumping with only a small amount of the actin appearing as single filaments. We have tried sonicating, gentle swirling for several hours, vortexing, and ammonium hydroxide.
As we understand it, the lyophilyzed product as purchased is G-actin that will polymerize to F-actin once in solution. However, we do not believe that the G-actin is going into solution, or at least that only a very small proportion of it is.
Any help would be greatly appreciated.
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
Are there companies which deal with buying and selling of 2nd hand TEMs? If so, I would appreciate contact names/emails/telephone numbers.
Thanks.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Has anybody had any experience of using the Nikon F90X with a Zeiss Axioskop? I'm not keen on the Contax 167MT that Zeiss want to sell me so I'm looking for an alternative. My main application is B/W photography of fluorescence staining.
If people are worried about file attachments, they shouldn't worry about it. Simply do NOT extract the file and if your mailer does it automatically, one can switch that option off somewhere in the options...usually just a flag switch. Usually I never extract attachments by default, because most of the time they are a waste of time and meant to be funny or something, and indeed it may slow your system down. Off the topic really, but just my two cents, which in our currency is worth sweet blow all, to calm down some some frantics on this list.
Democratically,
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
A simple answer to your question is a bit beyond the capability of this list server. However, we have several consultants on staff who can help you with your specific application. Please either visit our website ( { {http://www.MME-Microscopy.com/education} ) or call our office.
Best regards,
At 02:34 PM 8/19/98 -0300, MATERIALS-CARIRI wrote:
} } } }
{excerpt} {smaller} I will like to know the full use and range in testing materials on an scanning electron microscope model no. JSM-35C.At present tests that can be carried out are Microstructure, Particle Morphology and Micrographs.
Sally Burns wrote: } } We are attempting to fix chick forebrain neurons for TEM. Several cells have been processed and embedded in Marglass. A graded series was not used to dehydrate or embed, the blocs had serious infiltration problems. } To solve this I suggested using a graded series (with EtOH) and Spurtol. They tried and the Spurtol etched the plates. The plates are poly-L-lysine coated tissue culture dishes made by Falcon (Becton Dickinson). } Since the Spurtol was a problem, they have been experimenting with } Marglass and a graded EtOH series. However, when they try this, the plates are again etched. } Any suggestions?? Change resin? Change plates? suggestions on infiltration timing would also be helpful } } Thanks... Sally } Sally Burns } Center for Electron Optics } B5 Pesticide Research Center } (517) 355-5004 } burnssal-at-pilot.msu.edu
Sally:
I embed cultured neurons or glia in an Epon substitute without any problems. While I use EMbed 812 from Electron Microscopy Sciences I'm sure an Epon substitute from another vendor would work. Perform fixation, osmication, rinse, dehydrate as usual. When you finish with 100% alcohol (or even 95%) then go to 100%:resin 2:1, then 1:2, then two changes of pure resin. I usually get compulsive and leave the tissue culture dish in a vacuum dessicator overnight to get all of the solvent out of the last change. Then polymerize as usual. NO propylene oxide! "Epon" is completely miscible with ethanol even with a bit of water left over, it just does not mix as quickly as with prop. oxide. "Epon" (and its components) reacts very slowly with tissue culture plastic, Spurr and its components react very quickly (I actually did the experiments!) E-mail me if you need more advice.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (732)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
For my recent project I have to section perfused and cryoprotected rat brains 40 microns and do immunohistochemistry on floating sections. Sometimes the sections have to wait a couple of weeks before I do the immuno on them. I usually keep them in PBS in the fridge but always worry that it may lose some of it's antigenicity. In what would you recommend the sections be stored? Thank you, Lilith ------------------------------------------------------- Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
RESEARCH ELECTRON MICROSCOPIST to manage institutional core EM Facility and carry out all duties related to daily operation of the facility, including maintaining and using electron microscopes, preparing specimens, data collection, advising researchers on EM approaches, ordering supplies, record keeping and billing, etc. Requires minimum of Bachelor's degree and several years experience in biological electron microscopy; up to Ph.D. level considered. Expertise in several of the following techniques required: thin sectioning, immunolabelling, cryo-EM, negative staining, metal shadowing, cryosectioning, freeze fracture/etch, rapid freezing and freeze-substitution, EM autoradiography, in situ hybridization. Salary is highly competitive and commensurate with experience. Send CV and names of three references to Dr. George Witman or Dr. Roger Craig, Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Ave. North, Worcester, MA 01655. Email: Roger.Craig-at-ummed.edu or George.Witman-at-ummed.edu. The University of Massachusetts Medical School is an equal opportunity employer.
Hi Bob, My experience with lyophilization is that unless precautions are taken with the protein (glycerinization or ammonium persulfate precipitation) that you will end up with a bunch of agglomerated, insoluble blobs that will never go into solution. You might try adding some NaCl (1% w/v) or ammonium persulfate (same) to see if this will enhance suspension of the molecules. Did you call the Sigma tech reps? With some enzymes (glucosyl transferases from bacteria), we have had good luck storing the enzyme solution under a layer of toluene. Lasts for months. Let us know what you find out.
} Does anyone know how to get commercially available actin to disperse as } F-actin? We have not had good luck getting a lyophilyzed powder of porcine } actin (Sigma) into solution as dispersed F-actin filaments. We usually get } quite a bit of clumping with only a small amount of the actin appearing as } single filaments. We have tried sonicating, gentle swirling for several } hours, vortexing, and ammonium hydroxide. } } As we understand it, the lyophilyzed product as purchased is G-actin that } will polymerize to F-actin once in solution. However, we do not believe } that the G-actin is going into solution, or at least that only a very small } proportion of it is.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
---------------------- Forwarded by Sylvie Verdon/CRYOLIFE on 08/20/98 01:34 PM ---------------------------
Sylvie Verdon 08/20/98 01:21 PM
To: Microscopy-at-MSA.Microscopy.Com cc:
We do see some dirt here at the Ag Research Center, so I asked someone who had made soil pH measurements. Her reply:
It's a 1:1 dilution.
We check the pH of the field soils by using 10 g soil:10 ml 0.01M CaCl2 in a 15-ml screw cap test tube. Shake well, spin & measure pH. You can use d. h20 as we used to before changing to the CaCl2.
lorraine
via Leonard R. Corwin Fort Dodge Animal Health Princeton, NJ 08543-0400
The following position is available in our Department. (Please note that all responses should be addressed to Human Resources at the address given in the announcement, and not to me.)
Dr Gillian M. Bond Department of Materials & Metallurgical Engineering New Mexico Tech
NEW MEXICO INSTITUTE OF MINING AND TECHNOLOGY seeks qualified applicants for the position of Departmental SEM/Electronics Technician with Materials and Metallurgical Engineering. Candidates should be capable of operating and maintaining scanning electron microscopes, (SEMs), as well as maintaining and repairing other items of equipment. The position will involve interaction with both faculty and students, and participation in current research activities. Experience in the operation and maintenance of SEMs and other electronic equipment is required. New Mexico Institute of Mining and Technology has a rural location in central New Mexico, with easy access to Albuquerque. The setting provides an excellent university environment, and the opportunity to work with very active research groups and high-caliber students. Interested persons should send a complete resume with details of prior experience. Submit application material to New Mexico Institute of Mining and Technology, Human Resources, 801 Leroy Pl, Wells Hall Box 92D Socorro, NM 87801. For information about New Mexico Tech, visit our web page http://www.nmt.edu/. E-mail applications NOT accepted. AAEOE
For the purposes of immunoEM, I would like to localize GFP (green fluorescent protein) at the EM level. I hope to find a source of anti-GFP antibody, preferably tagged already with 10 nm gold particles. I've seen commercial sources of polyclonal anti-GFP. Does anyone know of a gold-linked version? David H. Hall Center for C. elegans Anatomy Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
Lisa D. Brown University of Connecticut Physiology and Neurobiology Department Electron Microscopy Laboratory Box U-131, Rm 129 Beach Hall Storrs, Ct 06269-2131 Tel. (860)486-2914 Fax. (860)486-1936
Responding to the message of {85256666.00601C0D.00-at-cryont.cryolife.com} from "verdon.sylvie-at-cryolife.com"-at-Sparc5.Microscopy.Com:
} Sylvie Verdon } 08/19/98 02:37 PM } } To: Microscopy-at-MSA.Microscopy.Com } cc: } Subject: } } Has anyone done a trichrome stain with GMA? We are looking for a trichrome } staining procedure on GMA embedded Plastic sections for viewing on a light } microscope. } } Thanks
Sylvie,
How about just a general plant stain for GMA embedded tissue, like toluidine blue or methylene blue?
Or do you want to stain for something more specific??
Gib
Gib Ahlstrand, Minnesota Micoscopy Society Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"Theory and practice are the same in theory, but different in practice."
I have used a variety of 35 mm cameras on several models of fluorescent microscope. My main concern with a camera such as the N90 is that unless the camera is parfocal to the eyepiece, the grain in its focusing screen makes composition and focusing difficult. A better solution would be a Nikon F3, F4, or F5. The normal viewing screen can be replaced with a clear "M" type screen and the prism replaced with a high magnification waist level finder. These cameras with appropriate screen will be easier to use than the N90. Also, these models of Nikon have mirror lockup switches, if you want to reduce the vibration induced by mirror slap. To install the Nikon in place of the Contax, I believe Zeiss sells suitable adaptor rings for their Axioscope.
Hope this helps. Brian Matsumoto Director of Microscopy Facility Neuroscience Research Institute University of California Santa Barbara, CA 93106
An understanding of the structure of foods and food products is essential to the efficient development of new processes or products. The future of the food industry is closely related to new developments in food microscopy.
This conference is an opportunity for those involved in food product development to hear the latest findings from key scientists in the field of food microscopy. The meeting will act as a forum to establish the needs of the food industry over the next decade, to discuss recent advances in food microstructural research and to evaluate new microscopical techniques.
This conference is aimed at food microscopists, food technologists and research scientists, particularly those involved in the development of new products or processes. Any company engaged in product development should be represented at the conference, to ensure that it is aware of the opportunities that advances in food microscopy can offer.
The aim of this conference is to:-
bring together food microscopists and food technologists to enable fundamental and applied research to be presented together;
highlight recent advances in food microstructural research;
provide a forum for discussion of microstructural studies relating to food;
identify the potential of new techniques.
Conference organisers: Mrs Kathy Groves and Dr Morag Saunders, Leatherhead Food RA; Dr Ashley Wilson, CCTR University of York.
There is still time for other papers to be submitted - please contact the conference organisers for further information.
KEYNOTE PAPER: Structure - the Only Thing that Matters? - Professor Peter Lillford, CBE, Unilever Research The challenge facing members of the food industry is to become architects of structure rather than processors of materials. This will need better application and novel approaches to microscopy.
Invited Paper: Cryogenic Environmental SEM of Ice Cream - an Introduction to the Principles of ESEM with Applications to Observations on Ice Cream - Brad Theil, Polymers and Colloids, Physics Department, University of= Cambridge
Food Microstructure using X-ray Projection Microscopy - John Judge and Peter Lillford
LUNCH
Chairman: David F. Lewis, SAC
The X-ray Microscope and its Applications to the Food Industry - Simon Burgess, Oxford Instruments
Probing Food Biopolymer Functionality with Atomic Force Microscopy (AFM) - Vic Morris, Institute of Food Research, Norwich (co-authors: A.P. Gunning, A.R. Kirby, A.R. Round, A. Mackie and P. Wilde)
The Environmental Scanning Electron Microscope - Speaker from Philips Electron Optics
Observations of Food Microstructure by Environmental Scanning Electron Microscopy - Debbie Stokes, Polymers and Colloids, Physics Department, University of Cambridge (co-author: A.M. Donald)
SESSION 2: APPLICATIONS OF MICROSCOPY TECHNIQUES IN FOOD RESEARCH
Chairman: Ashley Wilson, CCTR University of York
F Invited Paper: Applications of Variable Vacuum in Food Microscopy - Roger Angold, RHM Technology
F FTIR Microscopy in Troubleshooting for New Product Development - Hilary Holgate, RSSL
F Using Image Analysis and Confocal Microscopy Combined to Measure Deformation in Starch Matrices - Jeremy Addler, RHM Technology
F The Microscopy of Food. ESEM and Confocal Microscopy - Complimentary Techniques - Anthony Robinson, Polymers and Colloids, Physics Department, University of Cambridge
F Invited Paper: The Use of Electron Microscopy in the Investigation of BSE - Michael Stack, Veterinary Laboratories Agency
LUNCH and opportunity to view trade exhibition
SESSION 3: MICROSCOPY OF FOOD PRODUCTS, PROCESSES AND INGREDIENTS
Chairman: Roger Angold, RHM
Invited Paper: Microscopy - the Art of Food Technology - Professor Anne-Marie Hermansson, The Swedish Institute for Food and Biotechnology
Changes in Endosperm Structure during the Production of Popped Grain - Mary Parker, Institute of Food Research, Norwich
Confocal Laser Scanning Microscopy of Dairy Products and Ingredients - Methodology and Some Applications - Mark Auty, Dairy Products Research Centre, Moorepark, Ireland
Poster Exhibition and Opportunity to view Trade Exhibition
SESSION 3 (continued): MICROSCOPY OF FOOD PRODUCTS, PROCESSES AND INGREDIENTS
Chairman: Professor Anne-Marie Hermansson, SIK
Invited Paper: Brian Brooker, Institute of Food Research
The Use of SEM, Confocal Microscopy and Instrumental and Sensory Techniques in the Study of Biscuit Texture - Michael Edwards, Campden and Chorleywood Food RA
Crystallising Carbohydrates - Use of the Microscope in Understanding Crystallising Structures in Heat-resistant Chocolate and Powdered Sorbitol - Kathy Groves, Leatherhead Food RA
F Microstructural Changes of Food Products Following Inclusion of Non-starch Polysaccharides and Fibre Supplements: Implications to Dietary Nutrition - Karen Linton, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth (co:authors: C. Brennan and P. Moura de Magallraes)
LUNCH
Chairman: Brian Brooker, IFR, Reading; Mrs Kathy Groves, Leatherhead Food RA
Ultrastructural and Textural Changes in Heat-processed Fruits - Morag Saunders, Leatherhead Food RA
Cereal Structure: its Relationship to Raw Material Quality and End-product Utilisation - Charles Brennan, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth (co-authors: K. Linton, Seale-Hayne Department of Agriculture and Food Studies, University of Plymouth; D. Griggs, Pauls Malt Ltd; I. Cantrell, Pauls Malt Ltd; P.R. Shewry, IACR - Long Ashton)
Invited Paper: Perspectives on Food and Microscopy - David Lewis, SAC
CLOSE=09
ADMINISTRATION
Registration and closing times will be advised with the acknowledgement of your booking=09
Lilith- I would suggest freezing the tissue, probably prior to sectioning. Infiltrate tissue with OCT for 30-60 min. then Freeze, wrap in alum. foil and /or parafilm, and store in freezer (-20 C). I have done this in the past with human cornea, and antigenicity lasted months to years for the collagen Abs we worked with. -Mike
On Thu, 20 Aug 1998, Barry, Lilith wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleagues, } } For my recent project I have to section perfused and cryoprotected rat } brains 40 microns and do immunohistochemistry on floating sections. } Sometimes the sections have to wait a couple of weeks before I do the } immuno on them. I usually keep them in PBS in the fridge but always worry } that it may lose some of it's antigenicity. In what would you recommend the } sections be stored? } Thank you, } Lilith } ------------------------------------------------------- } Lilith Ohannessian-Barry } National Research Council } Institute of Biological Sciences } CANADA } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca } }
Fellow listmembers, Hope the day is going well for everyone. I am looking for some help. The department here (Dept. of Anatomy, National University of Ireland, Galway) is trying to set up a tissue culture facility and, although we have been lucky enough to get some grant money, we are having quite a bit of 'fun' trying to locate some equipment that we need at a price we can afford. Can anyone let me know of a good basic model of an inverted microscope that is relatively cheap and can be used for routine examination of cell cultures? We don't need any fluroescence or photographic capabilities, just the basic nuts and bolts microscope. Or, if anyone has a microscope that feels a bit of wanderlust and would like to live out its retirement in Ireland, we can offer a loving home!
I suppose emails directly to me would keep the list free from extraneous mails.
Since all the color correction, etc. has been done prior to the camera system, you should have no problem putting a Nikon camera on a Zeiss microscope. As a matter of fact, I think that Dr. Savile Bradbury (retired, Oxford), frequently used Nikon cameras on other types of scopes.
One place you may have to make a slight adjustment is on the parfocality (making sure that the microscope and the camera are both in focus simultaneously). One way to test is to set the microscope up for regular Koehler illumination (Brighfield) with the lowest mag objective (ex: 4x)using a thin, well-stained specimen. Mount the empty camera. (Zeiss has info on T-mounts which fit various types of cameras. As I remember, the Nikon's are usually bayonet mounted). Push/Pull the bar on the phototube so that all of the light is going to the camera. Open the camera back and place a small piece of diffuse at the film plane. (A small piece of Fresnel lens also works well). Adjust the height of the phototube until that image is perfectly in focus when the image is in focus for the microscope. From that point on, focusing the microscope should also ensure accurate focus at the film plane, for all magnifications equal to or greater than the objective you used.
For a more detailed discussion as to why all this works, I encourage you to see our book "Optimizing Light Microscopy for Biological and Clinical Laboratories". Details are available at our website: { {http://www.MME-Microscopy.com/education} .
1) Darkfield illumination is interesting because it shows many of the minute details. The preps need to be extremely clean, however, to avoid distraction from dust, etc.
2) Rheinberg illumination is a derivation. We have sets of photographic filters available for minimal cost. Please contact me off line for further information.
3) Hoffman Modulation Contrast might also work, although I have never used it in this particular application.
Details of all these techniques are in "Optimizing Light Microscopy for Biological and Clinical Laboratories" and details for the book are on our website: { {http://www.MME-Microscopy.com/education}
I do have one other contact who is a world-class expert on mites: Mike Huben. I will have to locate his information and send it to you.
Best regards,
At 03:06 PM 8/20/98 MST/MDT, Dr. Mark W. Lund wrote:
} From: Gary M. Easton {geaston-at-ibm.net} To: MSA {microscopy-at-sparc5.microscopy.com} Date: Friday, August 21, 1998 3:24 PMSubject: FREE KEVEX XRAY ANALYZER Hi all, !! FREE FREE FREE !! (I need the warehouse space!) Free to a good caring home - KEVEX Analyst 8000 EDS system(no detector). System is(as far as I've been told) operational. Includes all manuals and SEM interface. You arrange for the packaging and transportation and its yours. FREE. Please reply via email or phone. Gary M. Easton, Pres. SCANNERS CORPORATION Third party SEM service 410-549-3800 x102 gary.easton-at-scannerscorp.com
Please respond directly to the student if you can help him with his question.
Nestor
Dear sir/madam
I have been attempting to submit a question on the internet, but am not having much luck getting it through, please could you forward this question to the correct person.
Name : Alec Higgs School Name : Drakensberg Primary Grade/Level : 6 City : Newcastle State : Kwazulu-Natal Zip : 2940 Country : Republic of South Africa E-Mail address : alech-at-a6.new.iscorltd.co.za
Question :
I would like to know if different races have different shapes in their actual hair strand, Eg. oval or flat or round. (I do not mean curly or straight hair)
Please let me know if this does exist and detail the different races and their different shapes of their hair strands.
unsubscribe Dr. Massimo Catalano Istituto per lo studio di nuovi materiali per l'elettronica del Consiglio Nazionale delle Ricerche CNR-IME Via Arnesano 73100 Lecce - ITALY tel: +39 832 322362 fax: +39 832 325299 email: catalano-at-osfime.unile.it {http://www.ime.le.cnr.it/} http://www.ime.le.cnr.it
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Colleagues } } Please respond directly to the student if you can help him } with his question. } } Nestor } } Dear sir/madam } } I have been attempting to submit a question on the internet, but am } not having much luck getting it through, please could you forward } this question to the correct person. } } Name : Alec Higgs } School Name : Drakensberg Primary } Grade/Level : 6 } City : Newcastle } State : Kwazulu-Natal } Zip : 2940 } Country : Republic of South Africa } E-Mail address : alech-at-a6.new.iscorltd.co.za } } Question : } } I would like to know if different races have different shapes in } their actual hair strand, Eg. oval or flat or round. (I do not mean } curly or straight hair) } } Please let me know if this does exist and detail the different races } and their different shapes of their hair strands. } } Thank you for your response in this regard. } } Alec Higgs
About twenty years ago, I was employed by a company called "The Aerospace Corporation". They had several experimental projects under development. One was "gunshot residue" analysis which is common today. Another was the study of different hair samples. It was hoped that different hair samples could be linked to a given suspect (This was before DNA analysis was available). At the very least, it was hoped to correlate a hair sample to the suspect's race.
In a test, the scientist collected several hair samples from diffrent employees and tested their theory.
The results proved to be embarassing. Out of the dozens of hair samples collected, they concluded that only two hair samples matched. The donors of the two hair samples were the same race or at least related. The reality was that one sample came from a lady of European descent (with undyed blond hair) while the other was a male of African heritage. The program was discontinued.
Oh well, at least the "gunshot residue program" proved sucessful.
Dust mites and other mites can be made more visible (general morphology) by mounting in media with low refracting indice.
Lots of recipes are published: gum acc. to Faure, acc. to Berlese, acc. to Andre, polyvinyl alcohol lacto-phenol, gum dammar dissolved in 2-propanol, gum acc. to Apathy etc... I'll be glad to send you the recipes/protocols/references...
Impregnation in Amman's lacto-phenol and subsequent mounting in glycerin jelly acc. to Kaiser is also a usable method for general morphology.
I have had good results on Demodex foliculorum and Sarcoptes scabiei (an exoparasite in rabbits) with these staining techniques:
1. Staining with Ziehl-Neelsen carbolfuchsine -----------------------------------------------------------------
* Fixation in boiling ethylalcohol 70% the animals died in a perfect extended condition * Flood the object with a few drops of Ziehl-Neelsen carbolfuchsine and warm very gently until steaming * blot very gently dry (without touching the animals!) and differentiate with liquid phenol or acid alcohol (1 ml hydrochloric acid 37% in 100 ml ethylalcohol 70%) * mount in a water based non-acid mountant, or * dehydrate, clear and mount in DPX (numount, cedax...whatever).
2. Staining with pyrogallol --------------------------------------
* Stain the objects in 1% pyrogallol in distilled water or ethylalcohol 70% for about 30 min. * Bring the objects in distilled water or ethylalcohol 70% and let stand in bright daylight for a few hours, the color develops slowly.
When the color is to light, repeat the staining step; when the color is to intense, differentiate in acid alcohol (1 ml hydrochloric acid 37% in 100 ml ethylalcohol 70%)
* mount in a water based non-acid mountant, or * dehydrate, clear and mount in DPX (numount, cedax...whatever).
Staining with a saturated solution of picric acid in clove oil, followed by rinsing in xylene or toluene and mounting in DPX can give sometimes good results: the animals are stained a nice yellow-brown.
It's sometimes possible to stain exoskeletons of mites with Mayers hemalum, Grenachers boraxcarmine or hydrochoric acid carmine.
Yvan Lindekens, Belgium.
---------- } Van: Dr. Mark W. Lund {lundm-at-physc3.byu.edu} } Aan: microscopy-at-sparc5.microscopy.com } CC: lundm-at-physc3.byu.edu } Onderwerp: Stain for dust mites in light microscopy } Datum: donderdag 20 augustus 1998 17:06 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Can anyone recommend a way to stain dust } mites to make them more visible in } a light microscope? } } best regards } mark } }
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We like to inform about the Workshop "Mikromanipulationstechniken in der Mikroskopie " ( Language is German )
{H3} Mittwoch, 18. November {/H3} {HR SIZE=13 WIDTH="100%"} {BR} {BR} {TABLE BORDER COLS=3 WIDTH="100%" BGCOLOR="#FFFFFF" } {TR} {TD} 9.00-15.00 {/TD} {TD} Paktische Übungen an eigenen Präparaten nach vorheriger Absprache {/TD} {TD} {/TD} {/TR} {/TABLE}
{H3} Tagungsort: {/H3} Die Vorträge finden im Konferenzsaal des Hauptgebäudes, die praktischen Übungen im Seminarraum und den Mikroskopielabors des Laborgebäudes Chemie des IPF statt. {H3} Posterbeiträge: {/H3} Im Rahmen des Workshops ist eine Poster-präsentation zu Entwicklungen und Anwendungen von Mikromanipulationstechniken vorgesehen. Zur Festlegung des endgültigen Progamms sollte die Anmeldung der Posterbeiträge, ebenso wie von Kurzvorträgen auf der Tagungsanmeldung bis zum 25.10.1998 erfolgen. {H3} Tagungsgebühren: {/H3} Die Teilnahmegebühren in Höhe von {BR} 350,- DM respektive von 170,- DM für Teilnehmer mit Poster- oder Vortrag (nicht eingeladen) bitten wir unter dem Stichwort „Mikromanipulation“ auf nachfolgend aufgeführte Bankverbindung zu überweisen: {BR} Kto: 0526287200 BLZ: 850 800 00 {BR} Dresdner Bank AG Dresden {H3} Anmeldefristen: {/H3} Um den Workshopcharakter der Veranstaltung zu wahren, ist die Teilnehmerzahl auf maximal 30 Teilnehmer begrenzt. Eine rechtzeitige Anmeldung spätestens bis zum {FONT COLOR="#FF0000"} 25.10.1998 {/FONT} ist unbedingt erforderlich. {H3} Unterbringung: {/H3} Für Zimmerreservierungen wenden Sie sich bitte direkt an die Adressen der der Anmeldung beigefügten Liste von Hotels in unmittelbarer Nachbarschaft zum Institut, zum Bahnhof und zur Innenstadt oder an die Zimmervermittlung „Welcome Tourist“. {BR} {BR} {CENTER} {TABLE BORDER COLS=1 WIDTH="100%" BACKGROUND="10280033.gif" } {TR} {TD} {CENTER} {B} {FONT COLOR="#FF0000"} {FONT SIZE=+3} Versäumen Sie nicht ein Highlight 1998 ! {/FONT} {/FONT} {/B} {/CENTER}
Dear Fellow-listers, We are using the chicken embryo chorioallontoic membrane (CAM) model to study the blood vessel formation and its response to various treatments. I would like to quantify the results by measuring length, density amd branching of blood vessels using image analysis. I wonder if any of you is doing this and would appreciate any experience with image analysis, such as algorithms, type of software best suitable etc. Thank you,
Sarka Lhotak
Hamilton Regional Cancer Centre McMaster University Hamilton, Ontario, Canada
Hi =20 I am trying to inverse the gray levels of an image using the function=20 IpAnam (visilog software). =20 I have used the following=20 IpAnam("imagein";"imageout";{255,0},{0,255},0,255); =20 but this does not inverse the original image.....does anyone know what= =20 the arguments should be? I am afraid I don't quite understand the=20 explanations given in the manual =20 Franck. =20
I am considering selling our H-9000 HREM. This is a serious HREM with better than 1.8 Angstroms resolution, and comes with a Gatan TV camera. It has been (and still is) under maintenance contract. Some general information is available at:
http://www.numis.nwu.edu/internet/hrem.html
and contact me for further information.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
ELECTRON MICROSCOPY TECHNICIAN The Integrated Microscopy Core, Department of Cell Biology, Baylor = College of Medicine has an immediate full-time opening for an electron = microscopy technician. The Integrated Microscopy Core is a busy, = state-of-the-art facility with 2 TEMs, deconvolution, laser scanning = confocal and 2 CCD-based fluorescent microscopes. The applicant should = have at least 1-2 years experience in all aspects of sample preparation = for biological TEM including fixation, embedding, ultrathin sectioning = and staining of tissue samples and cell monolayers. The applicant = should have darkroom experience and experience in the operation of TEMs. = Immunolabelling experience is desirable but not essential. Other = duties include preparation of solutions, embedding media and the = maintaining of records. The position offers opportunities for training = in advanced light microscopy techniques, including fluorescence, laser = scanning confocal and deconvolution microscopy. The position requires a = minimum of a Bachelors degree and will start as=20
a Lab Technician II; salary range is low 20's, commensurate with = experience, and includes the standard Baylor benefits package.
Send CV and letter of research/technical interests to:
Hank Adams Laboratory Manager Integrated Microscopy Core Department of Cell Biology Baylor College of Medicine One Baylor Plaza Houston, TX 77030 Email submissions to: hpadams-at-bcm.tmc.edu Fax submissions to: 713 790 0545
Baylor College of Medicine is an Equal Opportunity, Affirmative Action = and Equal Access Employer.
I am about to compile a table of units of measure including SI, other metric, and customary units, with emphasis of those units used in biomedical sciences. The goal of this work is not just to enumerate all units (which is barely possible) but to list only the terminal symbols used as units in algebraical expressions of units. These unit atoms are associated with a precise semantics expressed by means of linear algebra.
Here I am interested in the unit symbols "low power field" (LPF) and "high power field" (HPF). These are commonly used in clinical medicine as a unit to measure the size of the reference system when counting material of interest in microsocpy. For example, erythrocytes, leucocytes in a unrine sediment can be reported as 10/HPF (10 per high power field).
Clearly, LPF and HPF are no exact units but rather qualifiers of semiquantitative observations. However, clinically, those observations are sometimes treated like hard numerical observations, including such uses as criteria for eligibility in clinical trials.
For inclusion into the table of units there are two options. Treat LPF and HPF independently as dimensionless units with value 1, which is the last resort option for all those difficult to compare units. However, this looses even the obvious relationship between LPF and HPF.
The other extreme is to treat LPF and HPF as proper volumes of fluid that overwiewed in the microsocope. This is probably hard to estimate and too variable.
Therefore, I could imagine treating the LPF and HPF as actual areas (measured in square-millimer) so that the per-LPF and per-HPF quantities would be convertible to areic numbers. While this is still incompareable to most other units in the system, it does at least establish a relationship between LPF and HPF.
I need the help of some expert in microscopy to give me an estimate of the typical view area in those magnifications typically called "low power" and "high power". It would also be of interest to know whether there are considerable subjective variations to those observations (i.e. what microscope is used? Monocular/binocular? Does the observer wear glasses? If so, does (s)he use apropriate oculars or normal oculars?) and to have an estimation of the error that results from those variations.
Any comment or advice is highly appreciated.
best regards -Gunther Schadow
Gunther Schadow ----------------------------------- http://aurora.rg.iupui.edu Regenstrief Institute for Health Care 1001 W 10th Street RG5, Indianapolis IN 46202, Phone: (317) 630 7960 schadow-at-aurora.rg.iupui.edu ---------------------- #include {usual/disclaimer}
Thank you very much for the reply on storing the floating sections question. There seem to be few options, all within the same idea. In fact I seem to have hard time deciding which one to use. If any of you interested, I will forward you the replies. Hope to be of help in the near future, Lilith ---------------------------------- Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
Can anybody tell me if the wax on the plant surfaces (aerial organs) does autofluoresce? Or, better off, send me to a reference. Sections of leek leaves show light green autofluorescence (filter BV-2A) at the surface of the epidermal cells, where the crystalline epicuticular wax is deposited. The cuticle may autofluoresce too, but I'm not sure. Thank you very much.
Camelia G.-A. Maier Postdoctoral Fellow The Samuel R. Noble Foundation Plant Biology Division Ardmore, Oklahoma
I will have to check on the exact procedure you are discussing but there are some very specific answers to your later questions:
1. The field of view for any objective can be calculated by dividing the field number by the total objective magnification.
a. The field number is usually engraved on the eyepiece and is typically something between 18 and 25. It refers to the diameter, in mm, across a small baffle or ridge which is placed anywhere between the mid line and lower 1/4 of the eyepiece. If you pull the eyepiece out, turn it upside down, then gently run a finger inside the barrel, you can usually feel it. In some types of eyepieces, (Huygen), it is mounted between the top and bottom (field) lens but you can usually still see it.
b. The total objective magnification is the magnification of the objective times the magnification of any other optics (tube lens, optivar or intermediate magnifier) up to but not including the eyepiece.
Ex:
For an 18mm field of view eyepiece and a standard 10x objective (most likely the one indicated by your LPF), the field of view would be 18mm/10. The resulting field of view is 1.8 mm or 1800 micrometers.
Ex II:
For a 20 mm field of view eyepiece and a 100x objective (most likely the one indicated by your HPF), the field of view would be 20 mm/100. The resulting field of view is 0.2mm or 200 micrometers.
If you used a 2x intermediate magnifier in this last example, it would drop the field of view to 100 micrometers.
2. This relationship holds irrespective of the brand of microscope used, of monocular/binocular, and with or without glasses.
For further information on these and related topics, might I suggest "Optimizing Light Microscopy for Biological and Clnical Laboratories". Details are available at our website. The book should be available locally through dealers who carry products from Structure Probe, Inc. If you cannot find it there, ordering information is also available on the website.
thanks for you quick and detailed answer! This helps a lot. When you say ``18 mm field'' or ``20 mm field'' in your example I assume that you speak of the diameter of a circular area, is that correct? It isn't by chance the radius?
} From what you say, I unfortunately have to conclude that the size of the so-called LPF and HPF depends on the type of eyepiece used by the observer. And the area seems to vary considerably as I calculated in the following table:
============================================= 18 FIELD NUMBER 25 --------------------------------------------- LPF (x10) 2.544 mm2 4.909 mm2 HPF (x100) 0.02545 mm2 0.04908 mm2 =============================================
This is a relative error of 63 % which is quite bad. When LPF and HPF are defined in terms of length (i.e. diameter) rather than area, I can press the error down to 32 % which is still way too high. It seems to me that really the only possible thing is to tell LPF from HPF by a factor of 100. So I could define LPF as 1 and HPF as 100. Any conversion to an actual area, let even volume, would be unreasonable due to this huge error that is inherent in those units.
} I will have to check on the exact procedure you are discussing but ...
So there is some last chance that LPF and HLP may be defined more exactly? Are there any recommendation by the Microscopy Society of America or some standards body concerning the use of LPF and HPF? Any standardizations or deprecations?
many thanks, -Gunther
Gunther Schadow ----------------------------------- http://aurora.rg.iupui.edu Regenstrief Institute for Health Care 1001 W 10th Street RG5, Indianapolis IN 46202, Phone: (317) 630 7960 schadow-at-aurora.rg.iupui.edu ---------------------- #include {usual/disclaimer}
Alec, I cannot answer your question...I am just awfully curious how you expect to define "race". Did you know that the differences within a so-called race, however you like to define it, is not significantly different from the differences between them. This is just the law of statistics that does not allow for such a differentiation, it is just in the prejudiced mind.
A fellow South African Quirina
} Dear sir/madam } } I have been attempting to submit a question on the internet, but am } not having much luck getting it through, please could you forward } this question to the correct person. } } Name : Alec Higgs } School Name : Drakensberg Primary } Grade/Level : 6 } City : Newcastle } State : Kwazulu-Natal } Zip : 2940 } Country : Republic of South Africa } E-Mail address : alech-at-a6.new.iscorltd.co.za } } Question : } } I would like to know if different races have different shapes in } their actual hair strand, Eg. oval or flat or round. (I do not mean } curly or straight hair) } } Please let me know if this does exist and detail the different races } and their different shapes of their hair strands. } } Thank you for your response in this regard. } } Alec Higgs
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
I'm trying to reach through e-mail Sales & Marketing Department of Energy Beam Sciences Inc. As so far without success. Actually, I'm not sure if it's the address or maybe our server's problem. Could anybody tell me if the address I'm using is the right one. It is: 75767,640-at-CMPUSERVE.COM.
Thanks in advance
Witold Zielinski Warsaw University of Technology Faculty of Materials Science and Eng. Narbutta 85 02 524 Warsaw Poland
A quick thank you to everyone that responded to the difficult task of cleaning oils... from powder metal samples. Many of the ideas have been tried in the past although with inconsistent results. The plasma etching method sounds promising although I have yet to try it as I am waiting for more samples. I am curious about the depth of cleaning while using this technique as the samples are usually a few cm thick .?. A quick run through will tell soon enough.
Thanks again. Wayne wengland-at-ortech.on.ca
XX
I just read your message and though I am no expert, a thought came to mind. Have you considered cleaning the metal powder in an oxygen plasma? I suspect that if it were to work you would probably have to clean as much of the oil off as possible by your current methods and try the plasma as the very last step. It might oxidize the surface of your metals, too. A question - when you heated the powder was that done in a vacuum or in air? We often heat our TEM samples in a (very good) vacuum to drive off hydrocarbons and have had good success.
I hope this is of some help.
xx
How about using an ESEM? It doesnt really care if there is some oil left, wash the stuff in a solvent and then put it in the ESEM. You can come and use ours if you want.
xx
Wayne, Do you have the capability to put the parts in a solvent reflux unit to provide a continuous wash over a 24 hr period? This should work especialy if it is set up to alloy the recondenced solvent to drip on the top of the part.
xx
Filter them onto a suitable membramne (i.e. Millipore) filter and flush it with a solvent for the oil. You'll have to choose a filter material that won't be dissolved by the solvent. If you precoat the filter with Au/Pd you shouldn't have any charging problems.
xx
Oil impregnated powdered metal is usually cleaned by extracting in petroleum ether. The process is to soak in ether, weigh the material, soak in ether and reweigh. Repeat the process until the weight does not change. Soak times may takes hours or even a few days.
xx
I was wondering what types of metals you were trying to clean? Also, when you heated the powder, did you heat it in a specific atmosphere of just an open air type furnace. And did you use any soaps or something that will 'capture' the oil molecules??
xx
Some Ideas for a very difficult problem. We have used a vacuum and clean in solvent in an ultrasonic cleaner. This process may require multiple repetitive steps.Running the parts through a degreaser if one is available may help. A degreaser that uses heat and vapor may be the way to go. If you have small parts a cold finger over an boiling acetone with a still the drips the condensed acetone on the part may be effective also. (TEM replica removal set is what I am trying to describe.) You may want to set a highly absorbent paper near where you want to look to help wick the oil way form the pores or crack(s). If you place the paper in a vacuum you may need to dry or out gas the paper first. If you are looking at oil impregnated parts such as bearings good luck. (I use Simple Green Cleaner, but this is not an endorsement of this product.) There are other water based solvents that work just as well or maybe better. Petroleum solvents work also but are harder to dispose of.
ASM in their metals handbook on metallography also has technique for cleaning samples. One old time engineer described this cleaning process as "worring over the part."
xx
Have you considered plasma cleaning? We market a system that was designed specifically for cleaning organic contamination from EM specimens. It utilizes a low energy, inductively coupled plasma to selectively remove the contamination without altering the specimen material. The process incorporates an oxygen plasma in which the disassociated oxygen created by the plasma chemically combines with the carbonaceous material. The resultant is a combination of CO, CO2 and H2O.
In our own laboratory when we have had small ball bearings and ball bearning raceways to be examined, and we don't want to use solvents, etc. out of fear of losing corrosion product or other materials from the analysis, we have used quite successfully our own SPI Plasma Prep II Plasma Etcher.
If you are not familiar with it, you can find it on our website URL http://www.2spi.com/catalog/instruments/etchers1.html
I toyed with the idea of making a public posting on the listserver but feared having it look "too commercial". However, if you should post any kind of summary, you certainly have my permission to post this information.
xx
In terms of actual metal powder used in powder metallurgical operations, when we receive them in the laboratory, they are often times containing something sorbed to their surfaces, but a quick exposure to an oxygen plasma within a few seconds etches away this layer resulting in a far better end result in the micrographs.
xx
I have been playing around with sovelnts (diff pump stack revival) Carbon tertracloride CCl4 fumes are brilliant. It boils at 76.54 deg Centegrade. Be sure that you work in a fumehood! Is used as a degraser at Cr and Ni coating facilitys.
xx
While I have not done SEM on powdered metals, I do have a bit of experience in cleaning the packing oils as I used to use powdered metals as reagents for high-temperature synthetic chemistry of intermetallics back in graduate school. I was extremely concerned about having no residual oil and as little surface oxidation as possible- also, several of the powdered metals I used were very reactive (in terms of reacting with air to form oxides). So, for your problem, it depends a lot on what the metal is (how fast it reacts with air) and how clean you want the surface to be. For extremely clean surfaces, I had friends in the physics department that would prepare a freshly cleaved surface from a bulk sample under the UHV conditions in their PES system. Techniques that I used involved repeatedly washing the powder in an appropriate solvent, using either filter paper on a buchner funnel or with special glassware having porous-glass filter-frits. For the very reactive metals (some of the lanthanides and occaisionally potassium or rubidium) I would work either on a schlenk line or in a nitrogen purged glove box using freshly distilled (extra-dry) solvents (usually pentanes, or hexanes or ether- depends on the oil- I'm not seure what is best for transmission fluid; I don't know what that is actually). For less reactive powders I could do a decent job with with solvents right out of the bottle working in air (in a fume hood)- then backfilling the container with dry N2 for storage. If the powder is very easily oxidized, you also have the problem of getting from an inert atmosphere to the SEM- for this you might get creative with a plastic bag that can be back-filled (or flushed several times) with dry nitrogen. There's lots of ways to do this kind of stuff on the cheap (if you aren't going to be doing it too often). You can find a lot of explanations and diagrams etc in a good inorganic (or organic for that matter) chemistry text book. The degree of care needed will be determined rather simply by the reactivity of the powder and the cleanliness that you require.
One disclamer is that several metal powders that are shipped under oil are done so as they react very violently with air or water- so be careful- (maybe you kno0w all this, but I don't want to be responsible) my advice is to check your library or local university bookstore for a good inorganic chemistry text so that you can know if their are potential hazards (MSDS can also be helpful- but they aren't always so well written).
Could someone responsible for submissions to the magazine Microscopy Today please reply to my personal address? A while back I was asked to contribute an article by someone whose e-mail address is no longer valid - I wish to find out if the contribution is still desired. Rob Palmer Director - Biofilm Imaging Facility CEB/UT
Witold Zielinski asked: } I'm trying to reach through e-mail Sales & Marketing Department of Energy Beam Sciences Inc. } As so far without success. Actually, I'm not sure if it's the address or maybe our server's } problem. Could anybody tell me if the address I'm using is the right one. It is: } 75767,640-at-CMPUSERVE.COM.
The Compuserve account has been closed. Our general e-mail address is ebs-at-ebsciences.com
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Our Epson FX-85 dot matrix printer which came with our System 4-plus EDS in 1986 finally died. Attempts to send the print command to later Epson printers (FX-86E) have not worked. Is there a fix or upgrade that has been successful by users from the RT-11 system? Thanks in advance.
} Can anybody tell me if the wax on the plant surfaces (aerial organs) does
} autofluoresce? Or, better off, send me to a reference. Sections of leek
} leaves show light green autofluorescence (filter BV-2A) at the surface of
} the epidermal cells, where the crystalline epicuticular wax is deposited.
} The cuticle may autofluoresce too, but I'm not sure.
} Thank you very much.
}
} Camelia G.-A. Maier
} Postdoctoral Fellow
} The Samuel R. Noble Foundation
} Plant Biology Division
} Ardmore, Oklahoma
}
}
Camelia,
One answer to your questions lays in the chemical structure of the wax. If it has conjugated bonds (alternating double and single) which allow mobility for pi bonding electrons, then it is likely to fluoresce. The more conjugation, the further the fluorescence will be to the red end of the spectrum.
One caution: chlorophyll is autofluorescent (green excitation, red emission) so it may be difficult to distinguish the wax if it is sitting on a leaf surface. You might try cross-sectioning the leaf and looking at the thin cross-section.
Re: defining LPF and HLP: yes. We have an expert Med Tech available but she is on vacation at the moment. I'll try to get you that info within the next week or so.
Re: the numbers 18 and 25. These are, indeed, the diameter across the open space defined by the baffle mentioned in the earlier posting.
I don't think defining these terms as arbitrary values of 1 and 100 does anyone any good. I think you need to be honest with what the numbers really represent. Since the American Optical Microstar 110 was the "industry standard" for so long. Unfortunately, those archives are packed up at the moment, but I think that the field number on the microscope was either 18 mm or 18.5mm. (The old American Optical line is now part of Leica so you might try calling their home office in Deerfield, IL).
concerning toxicity and waste disposal of fixatives, resins and other EM reagents I am looking for the print "Safety Chart, Chemicals in Electron Microscopy" from EMscope Laboratories Ltd., Kingsnorth Industrial Estate, Ashford, Kent. Has anybody the full adress or faxnumber of EMscope or could sent me a copy of the print? Furthermore I would be interested to know about the national guidelines for EM waste disposal, e.g. in the US and UK. Any comments are welcome!
Steven E. Slap, Vice-President P.O. Box 468 11 Bowles Road Agawam, MA 01001-0468 USA Phone: (413)786-9322 Phone:(800)992-9037 Fax: (413)789-2786
Click here to send mail: ebs-at-ebsciences.com ^^^^^^^^^^^^^^^^^^This may be what you're looking for
Tungsten filaments, Denka LaB6 and TFE cathodes; laboratory microwave processors; JB-4 microtome, triangular and Ralph knifemakers; Vibratome and MicroCut vibrating blade microtomes; Polaron sputter coaters, carbon coaters, critical point dryers, plasma ashers, vacuum evaporators and SEM cryo-preparation system; x-ray microanalysis standards; EM and histology embedding kits; EM automatic film processor.
regards -Gunther
Gunther Schadow ----------------------------------- http://aurora.rg.iupui.edu Regenstrief Institute for Health Care 1001 W 10th Street RG5, Indianapolis IN 46202, Phone: (317) 630 7960 schadow-at-aurora.rg.iupui.edu ---------------------- #include {usual/disclaimer}
Email, WWW sites, Phone, Product information, and address for hundreds of vendors are listed in the Directory of Microscopy Vendors at http://www.mwrn.com/vendors/directory.htm
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Thanks
Susanne
At 07:23 AM 8/25/98 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi Camelia, my experience is that epicuticular waxes do fluoresce but... Are the sections you're working on fixed? Glutaraldehyde in tissues that are resin embedded will result in a strong greenish fluorescence that is actually really welcome in some anatomical studies. You would have to look at some fresh tissue as a control. John
} } Hello all } } Can anybody tell me if the wax on the plant surfaces (aerial organs) does } autofluoresce? Or, better off, send me to a reference. Sections of leek } leaves show light green autofluorescence (filter BV-2A) at the surface of } the epidermal cells, where the crystalline epicuticular wax is deposited. } The cuticle may autofluoresce too, but I'm not sure. } Thank you very much. } } Camelia G.-A. Maier } Postdoctoral Fellow } The Samuel R. Noble Foundation } Plant Biology Division } Ardmore, Oklahoma
________________________ C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
We are a world-wide mineral company providing both natural and synthetic = mineral products to a number of industries including paper, plastic, = refractory, steel and food.
Title of Posted Position : Analytical Investigator Location: Easton, PA Division: Specialty Minerals, Inc. Department: Analytical Services
Typical Duties: Under minimal supervision, The Analytical Investigator (Microscopy) is = responsible for performing the chemical and microscopy investigations of = samples, maintaining technical mastery and awareness of the = state-of-the-art for areas of responsibility, performing administrative = duties, and actively assisting the Analytical Services Group achieve its = Objectives and fulfill its Mission. Primary duty is customer oriented = problem solving through the use of optical and electron microscopy and = microchemical analysis in a team environment.
Qualifications: Incumbent should possess a Bachelor's Degree in the physical sciences or = engineering, preferably in chemistry or mineralogy and have a minimum of = six years of chemical analysis or microscopy experience, preferably in a = service environment. The Analytical Investigator performs work of a = varied nature and is responsible for making some of and implementing = most of the decisions relevant to the areas of responsibility. Must have = excellent oral and written communication skills, computer skills, team = skills, ability to interact with a variety of people, and the ability to = operate analytical instrumentation and to manipulate equipment and = materials weighing 30-50 pounds. Previous analytical and LIMS experience = desireable.
All inquiries should be directed to the Human Resources Department:
Gary Duckwall, HR Manager Specialty Minerals, Inc. 640 N. 13th Street Easton, PA 18042
{!DOCTYPE HTML PUBLIC "-//W3C//DTD W3 HTML//EN"} {HTML} {HEAD}
{META content=3Dtext/html;charset=3Diso-8859-1 = http-equiv=3DContent-Type} {META content=3D'"MSHTML 4.72.3110.7"' name=3DGENERATOR} {/HEAD} {BODY bgColor=3D#ffffff} {DIV} {FONT color=3D#000000 size=3D2} We are a world-wide mineral company = providing=20 both natural and synthetic mineral products to a number of industries = including=20 paper, plastic, refractory, steel and food. {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} {/FONT} {/DIV} {DIV} {FONT color=3D#000000 size=3D2} Title of Posted Position=20 : = Analytical=20 Investigator {/FONT} {/DIV} {DIV} {FONT=20 size=3D2} Location: &= nbsp; &n= bsp; &nb= sp;=20 Easton, PA {/FONT} {/DIV} {DIV} {FONT=20 size=3D2} Division: &= nbsp; &n= bsp; &nb= sp; =20 Specialty Minerals, Inc. {/FONT} {/DIV} {DIV} {FONT=20 size=3D2} Department: = ; = =20 Analytical Services {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Typical Duties: {/FONT} {/DIV} {DIV} {FONT size=3D2} Under minimal supervision, The Analytical = Investigator=20 (Microscopy) is responsible for performing the chemical and microscopy=20 investigations of samples, maintaining technical mastery and awareness = of the=20 state-of-the-art for areas of responsibility, performing administrative = duties,=20 and actively assisting the Analytical Services Group achieve its = Objectives and=20 fulfill its Mission. Primary duty is customer oriented problem solving = through=20 the use of optical and electron microscopy and microchemical analysis in = a team=20 environment. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Qualifications: {/FONT} {/DIV} {DIV} {FONT size=3D2} Incumbent should possess a Bachelor's Degree in the = physical=20 sciences or engineering, preferably in chemistry or mineralogy and have = a=20 minimum of six years of chemical analysis or microscopy experience, = preferably=20 in a service environment. The Analytical Investigator performs work of a = varied=20 nature and is responsible for making some of and implementing most of = the=20 decisions relevant to the areas of responsibility. Must have excellent = oral and=20 written communication skills, computer skills, team skills, ability to = interact=20 with a variety of people, and the ability to operate analytical = instrumentation=20 and to manipulate equipment and materials weighing 30-50 pounds. = Previous=20 analytical and LIMS experience desireable. {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} All inquiries should be directed to the Human = Resources=20 Department: {/FONT} {/DIV} {DIV} {FONT size=3D2} {/FONT} {/DIV} {DIV} {FONT size=3D2} Gary Duckwall, HR Manager {/FONT} {/DIV} {DIV} {FONT size=3D2} Specialty Minerals, Inc. {/FONT} {/DIV} {DIV} {FONT size=3D2} 640 N. 13th Street {/FONT} {/DIV} {DIV} {FONT size=3D2} Easton, PA 18042 {/FONT} {/DIV} {/BODY} {/HTML}
I am at the University of Massachusetts Lowell. I have a VG ESCALAB II. The hardcopy output of the PDP11/83 is connected to an (obsolete) HP 7550A multipen plotter. After pen retrieval the pen comes down touches the paper, lifts back up and goes through its routine, for example, drawing the curve, but it is not in contact with the paper so it does not plot. The HP instructional book does not discuss this issue. Is it a software or a harware problem?? I need some direction. Thanks Dan Oblas
Hi there folks, I think the problem may be hardware if it does that with every color pen. Now there were different length pens at one tie also. Are you sure of what you have loaded? Just a thought out of a foggy ex-hp resellers engineers brain! Ed Sharpe
Microscopy Vendors Database at http://www.kaker.com is currently accessible only under IP address http://207.137.96.185/mvd/vendors.html, because we are in process of replacing current ISP with new one.
I am looking for THIN-pointed (not welded-pointed) filaments for our Siemens Elmiskop 102 electron microscope. Does anybody have some information about this article ?
Thanks in advance
Philippe Drouillon Solvay Research and Technology Electron Microscopy and Image Analysis Rue de Ransbeek, 310 B-1120 Brussels (Belgium) phone : (00 32) 2 264 24 47 fax : (00 32) 2 264 20 55 mailto:philippe.drouillon-at-solvay.com
------------------------------------------------------ Dr. B=E9la P=E9cz Research Institute for Technical Physics and Matl. Sci. H-1525 Budapest, POBox 49 Hungary phone: 36-1-395-9240 fax: 36-1-395-9284 E-Mail: pecz-at-mfa.kfki.hu ------------------------------------------------------
Philippe Drouillon asked: } I am looking for THIN-pointed (not welded-pointed) filaments for our } Siemens Elmiskop 102 electron microscope. } Does anybody have some information about this article ?
We (Energy Beam Sciences) have manufactured a range of pointed filaments for almost 30 years. There are several styles available, and they can be mounted on any filament base. Please contact me back-channel for additional information, or visit our website (address in my .sig, below).
I am looking for a distributer in Germany/Europe for antibodies from Miles Laboratories Inc. I even cant get a valid (www)-address of the mother company.
i am trying to determine the grain size of aisi 1050 that has been spheroidized is there an etch that would let me see the grains and not the carbon phase?
I am looking for a pan keratin antibody that works at the EM level on cultured keratinocytes that have been fixed in Zambonis (para-picric acid), and embedded in LRWhite.
I need to lable the cyto-skeleton and also our protien of interest. Does anyone know of one that works?
Can anyone help me with an address, web-site, or phone number for RMC in the US? We are looking at purchasing an ultramicrotome with cryo-adaptor. Anyone have any preferences between RMC's and Leica's models? Thanks Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219 (214) 559 7744
Professional electron microscopist with a minimum of 5 years experience to manage the daily activities of the EM Facility at the University of Missouri-Kansas City School of Dentistry. Duties would include active participation in ongoing research, troubleshooting, scheduling, and training on facility instruments: Philips CM12 STEM with new Princeton Gamma Tech EDS system; Philips 515 SEM with new Princeton Gamma Tech EDS system and Robinson backscatter electron detector; and recently-installed Philips Environmental SEM equipped with Schottky field emission source, gaseous electron detector and new Princeton Gamma Tech EDS system. Applicant should have operational and maintenance experience with transmission and scanning electron microscopes. Additionally knowledge of energy dispersive spectroscopy and specimen preparatory techniques is required. The position is affiliated with the Department of Oral Biology at UMKC School of Dentistry. Salary is commensurate with level of education and experience. Please respond by E mail to "eickd-at-umkc.edu" with resume.
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} Posted-Date: Tue, 25 Aug 1998 16:29:53 +0200 } X-Sender: neubohn-at-mendel.ipk-gatersleben.de } Date: Tue, 25 Aug 1998 16:29:53 +0200 } To: microscopy-at-sparc5.microscopy.com } From: Birgit Neubohn {neubohn-at-IPK-Gatersleben.de} } Subject: toxicity of EM reagents } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Given Birgit requests guidelines and comments, this may be of interest. It was originally sent to MICROSCOPY TODAY, but was not published, presumably because the topic was viewed as parochial. ***** Uranium compounds, especially uranyl acetate, have been widely and routinely used as transmission electron microscopy (TEM) contrast stains for biological materials since 1958 (1,2). Those who do TEM of biologicals use small quantities of uranyl acetate, nitrate, formate, sulfate and perhaps other uranuim compounds almost daily and therefore keep inventories of these salts and their solutions.
In the 1980's growing concerns about medical and research wastes entering regional dump sites prompted state radiation officials in Oregon to begin tightening the regulations for monitoring and controlling all radioactive substances, including the uranium compounds commonly used in processing biological specimens for TEM. Oregon has come to be a state with high levels of awareness about environmental and safety issues and is often at the forefront of regulatory and management trends in these areas. What happens in Oregon may, therefore, frequently serve as both a harbinger of changing attitudes and a model for standards which are evolving elsewhere. Messages seen in electronic news groups and conversations with colleagues suggest that other states may have also addressed, or may plan to address, the purchase, storage, distribution, use, and disposal of uranium compounds used for staining biological preparations for TEM.
Radiation-producing machines and radioisotopes used and stored at locations under State of Oregon jurisdiction are subject to provisions of the Oregon Rules for Control of Radiation, set forth by the Radiation Protection Services, Health Division, Oregon State Department of Human Resources. In addition, radioactive material transport, storage, and disposal must comply with rules issued by the Oregon Department of Energy, Oregon Department of Environmental Quality, and applicable federal agencies such as the U.S. Nuclear Regulatory Commission and U.S. Environmental Protection Agency. Regulations covering naturally occurring radioactive materials (NORM), a class of materials which includes uranium compounds used as stains in EM, have been in place in Oregon for several years (3). When these regulations were established, Oregon State University (OSU) undertook regulatory investigations to determine if use of small quantities of NORM could, or should, fall within the guidelines for exemption allowed under these regulations.
In 1992 the investigations were completed and a process to include uranium compounds used as stains for electron microscopy in the state's radiation regulatory and safety program was begun. Hence, it was at this time that our use of uranium-based compounds was first called into question. There are may isotopes of uranium. Depending on what mix of these one may be dealing with, various levels of alpha, beta, and gamma radiation can result from their radioactive decay.
On the OSU campus, the radiation safety program is managed by a radiation safety officer responsible to university administration and a University Radiation Safety Committee. The radiation safety officer supervises radiation safety training programs, use, storage, disposal and licensing matters, and coordinates functions of a faculty committee which establishes local policy and reviews safety and compliance matters.
Prior to 1992, neither facility magagement nor our radiation safety officials felt the small amount ( {25 grams with activity {10 microcuries) of NORM uranium compounds on hand in the facility, and their limited methods of use, posed sufficient hazard to warrant regulatory action.
Campus radiation safety personnel have always been understanding, knowledgeable, and non- adversarial in carrying out their responsibilities, but as the state mandates for stricter control were imposed, neither facility management nor the radiation safety officer were sure how best to proceed, with minimal adverse impact, toward compliance. The EM Facility at OSU is a service laboratory: several dozen students, technicians, and faculty use the facility and its supplies, and uranium-stained tissue embedments and grids are typically dispersed into the possession of facility clients. If materials that contained uranium were used by large numbers of people or removed from the "authorized" facility, could we allow these use and dispersal practices and still comply with the spirit and intent of the stricter regulations? The concerns went beyond possible health effects from radioactivity to include possible health effects related to more conventional chemical toxicity issues and the possibilities for contamination effects where inadvertent disperal might result in inaccuracies in low-level radiation monitoring activities.
We began by discussing and arriving at a mutual understanding of the regulatory needs and concerns and the complications that altered protocols and compliance-management would entail on both the EM facility and the radiation safety program. We next undertook to quantify the hazard potential. Our small stock of NORM uranium compounds in powder form were identified, inventoried, weighed and surveyed for radiation levels within their containers (bottles) and exterior to the sealed containers. Two dilute uranyl acetate solutions were likewise identified, inventoried, and quantified. In addition, we processed an assortment of plant, animal, and microbial specimens through standard protocols and submitted for analysis samples of the tissues, pellets, and the used and unused processing solutions before and after uranyl compound staining. We also provided samples of the plastic embedded tissues and pellets, as well as sections on copper TEM grids cut from these embeddments.
One interesting complication in quantifying the hazard arose early in the process. Every other campus user of radioisotopes was using these materials as tracers. These users were concerned with specific activity. As a consequence, their inventories were quantified and managed in protocols in terms of microcuries. For electron microscopy, we were concerned only with the electron scattering potential of the uranium atoms. We quantified our inventories and formulated our solutions on the basis of weights and volumes. A calculation of 0.33 microcurie/gram of U-238 was made to convert uranium compound supplies from weight to specific activity units for inclusion under the hazard assessment required by the stricter regulations.
Radiation levels (millirem/hr) were measured from open and closed containers of NORM radiation compound powders and solutions. Specific activity values associated with these measurements were then calculated from the specific activity of depleted uranium and the weight percent of uranium in the compounds. Working strength uranyl acetate solutions, nominally 1% w/v, gave specific activity measurments below 1x10-3 microcurie.
A variation of neutron activation analysis, gamma-ray spectroscopy using a germanium/ lithium detector, was required to detect uranium in processed tissues and pellets, where uranium levels in the 0.8 to 0.9 ppm range were reported. To assure a high level of accuracy, the analysis equipment was calibrated a number of times, and the results from samples submitted in December 1993 were not obtained until May 1994. The presence of uranium in stained materials on grids was below the detection limit of the analytical equipment and procedures used.
The final result of the quantitative and regulatory processes produced the development and implementation of a policy by which the Radiation Safety Office and EM Facility agreed to cooperate to enforce defined procedures. These are the central features of this policy.
A) Persons using uranium compounds in the EM Facility take a four hour radiation safety training class from our campus radiation safety officer. They then receive authorization to use NORM uranium compounds as stains in the facility. Individuals who want to purchase and maintain their own stocks of uranium-based staining compounds at other locations must obtain a radiation use authorization (RUA) permit for those locations from the campus Radiation Safety Office.
B) After use authorization is granted, users may obtain and manage their own stocks of uranium compounds or may use uranium stain solutions in the EM Facility. Uranium salts and stain solutions provided for use by and in the EM Facility may not be taken to other locations.
C) Persons using uranium-based stains in the facility are personally responsible for complying with all requirements for use, clean-up, in-lab disposal, and radiation monitoring. The EM Facility provides radiation safety and monitoring, clean-up, and disposal materials for client use.
D) The EM Facility manager maintains and documents the status of the uranium compound and radiation safety materials inventory, monitors for indications of radiation contamination or unsafe procedures, and attends to the safe and timely disposal of accumulating radioactive wastes.
E) Specimens which have been stained with any uranium compound(s) for EM and subsequently either solvent-rinsed, plastic embedded, or put on grids may be removed from the EM Facility for archiving or futher processing at other locations as long as uranium concentration gives a specific activity below 0.05 microcurie/gram.
This policy took effect 01 January 1994. Despite a small burden of additional managerial tasks and the minor inconveniences of more record-keeping chores, the policy has, to date, worked well.
Two additional points should be made. Authorization of NORM use in Oregon, and presumably elsewhere, is rather complex. Under Oregon regulations there is still some exemptions allowed for specific NORM uses, materials, concentrations, and amounts. Users of uranium compounds should investigate their specific situation, giving consideration to ALL uses of NORM at their organizations, not only the use of uranium for EM stains.
Finally, it should be noted that a particular problem is associated with the disposal of uranyl nitrate waste. This compound is a radioactive substance, a nitrate, and an oxidizer, and therefore is classified as a mixed radioactive and chemcial hazard.
1) Watson, M.L. (1958). J. Biophy. Biochem. Cytol. 4, 475. 2) Swift, H., Rasch, E. (1958). Sci. Inst. News 3, 1. 3) Radiation Safety Manual, Oregon State University
Al Soeldner Lab Manager EM Facility Oregon State University Corvallis, OR 97331-2902
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I have a user who wants to try to capture a specific virus from a prep contaminated with TMV. I believe there are methods involving placing a layer of antibody against a coat protein from the virus you want to capture on a grid. Then you float the grid on a droplet of the virus prep, wash off any unbound particles and negative stain. Hopefully this would leave only the desired virus on the grid. Does anyone use this technique and/or have a more detailed procedure?
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
by imo15.mx.aol.com (IMOv14_b1.1) id NRBOa20769 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 26 Aug 1998 17:47:56 -0400 (EDT) Message-ID: {b475c5c4.35e4828d-at-aol.com}
need micro manipulation apparatus for use on light microscope. please contact ed sharpe thanks
We have a Polaron diode sputter coater we use exclusively for gold coating for conventional 20 kV tungsten emitter SEM.
Lately the coatings being applied seem comparatively thin - instead of a shiny metallic gold coloured coat we get a semitransparent dark gray coat.
But we are coating using the same parameters as ever : 50 mA -at- 1.4 kV with 0.2 torr of argon.
This occurs even with plain glass specimens which should not be contributing any unwanted vapours.
I have checked out the pumping system and the argon gas bottle is labelled "high purity argon" and is quite new.
I would welcome suggestions as to the source of our problem.
Mel Dickson
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
} Dear sir/madam } } I have been attempting to submit a question on the internet, but am } not having much luck getting it through, please could you forward } this question to the correct person.
} } Name : Alec Higgs } School Name : Drakensberg Primary } Grade/Level : 6 } City : Newcastle } State : Kwazulu-Natal } Zip : 2940 } Country : Republic of South Africa } E-Mail address : alech-at-a6.new.iscorltd.co.za
} } Question : } } I would like to know if different races have different shapes in } their actual hair strand, Eg. oval or flat or round. (I do not mean } curly or straight hair) } } Please let me know if this does exist and detail the different races } and their different shapes of their hair strands. } } Thank you for your response in this regard. } } Alec Higgs
My appologies for the lateness in this reply, I have been out of the country. As an old and still practicing classical hair comparison person my views may be seen by many as biased.
The simple answer is that there has been found a relationship between cross sectional shape and racial characteristics. Flat being associated with the Negroid race, oval with the Caucasion race, and round with Mongoloid.
To quote from "MICROSCOPY OF HAIR, A Practical Guide and Manual" put out by the FBI:
"A human hair may be classified according to its racial characteristics as being Caucasian, Negrod or Mongoloid origin. In some instances, the racial characteristics exhibitied by the hair specimine may not be clearly defined indicting the sounce of the particular hair may be of mixed racial origin."
and
"Again, it is pointed out thet even when racial characteristics are not clearly defined, it is significant when these characteristics are consistent between the hair in questiion and the hair of known origin."
Okay, probably more than you wanted to know. Good luck.
Shalom from Jerusalem, Azriel +++++++++++++++++++++++++++++++++++++++++++++++++++++ Major Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem, ISRAEL
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
What lies behind us and what lies before us are tiny matters compared to what lies within us. Ralph Waldo Emerson ++++++++++++++++++++++++++++++++++++++++++++++++++++
I am interested in micro-manipulation (e.g. handling, applying external forces, injection................) of structural elements. Any information on the issues mentioned below would be of interest:
* instrumentations/instrument configurations * colleagues with expertise in this area * workshops, courses, conferences * manufacturers of instrumentation * handbooks, literature references
Please reply to:
Marcel Paques Wagenigen Centre for Food Sciences P.O.Box 20, 6710 BA Ede telephone: (31) 318 659690 telefax (31) 318 650400 email: paques-at-nizo.nl
I am posting this query on behalf of a colleague. He wishes to locate a supplier of microscope slides which have fluorescent beads mounted on them. He knows of suppliers which will supply him with the actual fluorescent beads but he wants them already mounted. He thought a company called "Phoenix" may be able to help him but he can't locate the company. The slides (with fluorescent beads attached) will be used to assist in optimising the performance of a Bio-Rad Confocal MRC1000. Any help locating these slides would be appreciated.
Thanks in advance,
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
Ed Sharpe asked about micromanipulators for light microscopes. Energy Beam Sciences distributes two lines of micromanipulation equipment. Information is available on-line at our web site (address in my .sig, below). Please contact me back-channel for additional information.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com ********************************
Does anyone have any suggested sources, recommendations, or comments regarding any software for tracking "cells" / motion analysis from microscopic images? Either live time, from video tape or a series of digital stills? Rather than trying to write software from scratch or as a series of macros it would be nice to find a ready to run package.
Aparently there used to be a package called "CellTrak" but a variety of web searches has resulted in nothing.
(Vendors should feel free to contact me as well.)
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
My first thought would be contaminated gas, either from a vacuum leak or a "bad" bottle of argon. The leak is more likely. With no Ar flow is your blank-off pressure the same low value as when working properly? I would also inspect the target/chamber for contamination of some sort. Another check.. If the pressure gauge is "off" you should notice a different current for the same voltage setting while sputtering.
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Hello to the coating experts:-
We have a Polaron diode sputter coater we use exclusively for gold coating for conventional 20 kV tungsten emitter SEM.
Lately the coatings being applied seem comparatively thin - instead of a shiny metallic gold coloured coat we get a semitransparent dark gray coat.
But we are coating using the same parameters as ever : 50 mA -at- 1.4 kV with 0.2 torr of argon.
This occurs even with plain glass specimens which should not be contributing any unwanted vapours.
I have checked out the pumping system and the argon gas bottle is labelled "high purity argon" and is quite new.
I would welcome suggestions as to the source of our problem.
Mel Dickson
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
id AA49470; Thu, 27 Aug 1998 08:08:05 -0400 Message-Id: {9808271208.AA49470-at-rose.muohio.edu} Received: from CASSERVER1/SpoolDir by casmail.muohio.edu (Mercury 1.32); 27 Aug 98 08:08:06 -5 Received: from SpoolDir by CASSERVER1 (Mercury 1.32); 27 Aug 98 08:07:43 -5 To: microscopy-at-Sparc5.Microscopy.Com, CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU
Looking for recomendations for a film scanner. The Leaf 45 is no longer in production. So far I have only found the Nikon LS-4500AF and the Polaroid Spirit Scan 45 Pro. Any others? Any recommendations?
Flatbeds are o.k. for 4X5's (we already have one) but would also like to be able to handle 35mm's as well (8x10 } 9x14 flatbed scanners just don't have the resolution for 35mm's).
Thanks again.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
Dear all, we found paracrystalline ER-structures in cells of the nectarie= s of plant flowers. Has anyone seen similar structures or knows about such things documented in t= he literature? Thanks for help Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=E4tsstrasse 25 Germany 33615 Bielefeld phone: 0521 1065592 fax: 0521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws
} I would like to know a way to grind and polish diamond samples for a SEM } study. What do you want to do for diamond samples. If you just observe the morphology of the samples by SEM, the samples needn't polish and grind. I ever used an agate motar to grind diamond, but the agate motar was worn out.
You may increase the number of a particular virus if the grid has been treated with antibody, but, it does not stop TMV from attaching to the grid. Why not treat the extract with TMV antibody and centrifuge it. You should be able to get rid of all TMV if correct amount of antibody is used.
Ann Fook Yang EM Unit Eastern Cereal and Oilseed Research Centre Agriculture and Agri-Food Canada 960 Carling Ave Central Experimental Farm Ottawa, Ontario Canada K1A 0C6
} } } Debby Sherman {sherman-at-btny.purdue.edu} 08/26/98 05:49pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a user who wants to try to capture a specific virus from a prep contaminated with TMV. I believe there are methods involving placing a layer of antibody against a coat protein from the virus you want to capture on a grid. Then you float the grid on a droplet of the virus prep, wash off any unbound particles and negative stain. Hopefully this would leave only the desired virus on the grid. Does anyone use this technique and/or have a more detailed procedure?
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Maybe I'm going blind. I've been digging through my Noran Voyager manuals and I can't seem to find KeV and take-off angle limits for the Proza quant. routine. Does anyone know the limits or can they direct me to the correct page(s)?
Thanks,
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 (978) 750-1717 crossman-at-osi.sylvania.com
I have a user who wants to try to capture a specific virus from a prep contaminated with TMV. I believe there are methods involving placing a layer of antibody against a coat protein from the virus you want to = capture on a grid. Then you float the grid on a droplet of the virus prep, wash off any unbound particles and negative stain. Hopefully this would leave only the desired virus on the grid. Does anyone use this technique and/or have a more detailed procedure?
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu=20 Purdue University =20 1057 Whistler Building West Lafayette, IN 47907-1057
The technique you descirbe is the "immunosorbent technique" and relies as you know on the fact that proteins tend to stick to the coated grids, especially a plastic such as formvar (which can be carbon coated to make it more stable- but then proteins do not stick as well). Antibodies appear to stick well to formvar-carbon and in my experience, once they are "on" its pretty well impossible to get them off!
This method will not completely get rid of your TMV contamination, that will stick to the grid as well (but will at least be a useful "standard" for magnification calibration!), however, the immunosorbent technique should enrich the number of specifically adsorbed particles which react with the antiserum. This is great for negative staining viruses when you only have a small volume of very dilute particles- so long as you have an antibody which you know reacts with the surface of the particle.
You will have to play with the conditions to get the optimum results with your anitbody, try a few different dilutions of it.
To start with, I suggest the following:
1. Float the grids on your antiserum- 10 min(all reactions carried out in humid chamber to prevent drying out) 2. Wash in PBS containing .1 per cent BSA 30 seconds 3. Float grids (antiserum coated side down!) on your virus preparation, for 5 minutes. 4. Blot of excess, float on PBS/BSA as above to wash. 5 Brief wash in deionised warter (optional) 5. Negative stain using your favourite stain UA, PTA etc.
best of luck!
Dr Timothy F. Booth Canadian Food Inspection Agency National Centre For Foreign Animal Disease Suite T2300 1015 Arlington St. Winnipeg Manitoba R3E 3M4 CANADA http://www.hc-sc.gc.ca/main/lcdc/web/bmb/fedlab_e.html#toc email tbooth-at-em.agr.ca Tel 204 789 2022 (office) Tel 204 789 2038 (lab) Fax 204 789 2038
You might want to take a look at the UMAX Powerlook 3000. Paper specs look pretty good and at $6995 (list), the price is less than the Polaroid.
http://www.umax.com/
Henk
At 08:07 AM 8/27/98 -0500, you wrote: {snip} } } Looking for recomendations for a film scanner. The Leaf 45 is no longer in } production. So far I have only found the Nikon LS-4500AF and the Polaroid Spirit Scan } 45 Pro. Any others? Any recommendations? } {snip} } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981 } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Another flatbed scanner which we considered a few years ago:
Dicomed 6300T: 35 mm to 4"x5", 12 bits/pixel, 6000 pixel CCD optical resolution (landscape mode ): 4233 dpi for 35 mm; 1270 dpi for 4"x5" Dicomed, 12270 Nicollet Ave., Burnsville, MN 55337, (612) 895-3000
} ------------------------------------------------- } Looking for recomendations for a film scanner. The Leaf 45 is no } longer in } production. So far I have only found the Nikon LS-4500AF and the Polaroid } Spirit Scan } 45 Pro. Any others? Any recommendations? } } Richard E. Edelmann, Ph.D. -------------------------------------------------
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello to the coating experts:- } } We have a Polaron diode sputter coater we use exclusively for gold coating } for conventional 20 kV tungsten emitter SEM. } } Lately the coatings being applied seem comparatively thin - instead of a } shiny metallic gold coloured coat we get a semitransparent dark gray coat. } } But we are coating using the same parameters as ever : 50 mA -at- 1.4 kV with } 0.2 torr of argon. } } This occurs even with plain glass specimens which should not be } contributing any unwanted vapours. } } I have checked out the pumping system and the argon gas bottle is labelled } "high purity argon" and is quite new. } } I would welcome suggestions as to the source of our problem. } } Mel Dickson } } } ***************************************************** } Mel Dickson, } Director. } Electron Microscope Unit, } University of New South Wales. } Sydney NSW 2052 Australia } } Phone (+612) 9385-6383 } Fax (+612) 9385-6400 } Website {http://srv.emunit.unsw.edu.au} } ***************************************************** } } Hi Mel,
Check for an air leak on your Argon input line particularly between the chamber and the leak valve. Black gold is always caused by air leaks on our sputter coater.
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
thanks for the info! I am trying to do things on a budget and am willing to accept old crusty equipment that even needs work done to it! I wish I had the budget for new equipment but sometimes we make do as best we can!
thanks though! Dear fellow microscopists,
Ed Sharpe asked about micromanipulators for light microscopes. Energy Beam Sciences distributes two lines of micromanipulation equipment. Information is available on-line at our web site (address in my .sig, below). Please contact me back-channel for additional information.
Best regards, Steven E. Slap, Vice-President
******************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com *
Dear Mel, If the operating parameters are the same, then the obvious culprits are the gold target getting thin or gone, or contamination of the gold target by vapours of some kind. Try cleaning the target with acetone or clean alcohol. You can even polish it lightly. You might also check all the high voltage contacts and the contact of the gold to the plate. Good luck. } } } Hello to the coating experts:- } } We have a Polaron diode sputter coater we use exclusively for gold coating } for conventional 20 kV tungsten emitter SEM. } } Lately the coatings being applied seem comparatively thin - instead of a } shiny metallic gold coloured coat we get a semitransparent dark gray coat. } } But we are coating using the same parameters as ever : 50 mA -at- 1.4 kV with } 0.2 torr of argon. } } This occurs even with plain glass specimens which should not be } contributing any unwanted vapours. } } I have checked out the pumping system and the argon gas bottle is labelled } "high purity argon" and is quite new. } } I would welcome suggestions as to the source of our problem. } } Mel Dickson } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Is there any chance that the Au target has worn through in spots? If so, it may be possible that sputtering is taking place with the underlying metal instead of or in addition to the Au.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML--Biology New Mexico State University Las Cruces, NM 88003
Flatbed scanners with transparency capability may offer the capability you need at much lower price than a 4 x 5 film scanner or drum scanner. Several models now costing a few $K have optical resolutions to } 2K and 14-bit grayscale. For a recent review, see Macworld October, 1998, pp77-81. If possible, try before you buy and look carefully at the control software. Scanning times can be longer than you can tolerate (How long does it actually take to scan each negative?), and manual scanner controls tend to be primitive or arcane (often both). Even 12-bit grayscale is usually sufficient for high-contrast negatives including TEM diffraction patterns if these are properly exposed. Manual exposure control would be useful for dealing with over/under-exposed film, but is seldom included.
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA
---------- From: Hendrik O. Colijn Sent: Thursday, August 27, 1998 4:23 PM To: edelmare-at-muohio.edu Cc: microscopy-at-Sparc5.Microscopy.Com Subject: Re: 4" x 5" film scanners?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Richard,
You might want to take a look at the UMAX Powerlook 3000. Paper specs look pretty good and at $6995 (list), the price is less than the Polaroid.
http://www.umax.com/
Henk
At 08:07 AM 8/27/98 -0500, you wrote: {snip} } } Looking for recomendations for a film scanner. The Leaf 45 is no longer in } production. So far I have only found the Nikon LS-4500AF and the Polaroid Spirit Scan } 45 Pro. Any others? Any recommendations? } {snip} } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981 } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
One way is to apply the diamond surface to a hot iron alloy flat - this has been done at the naval research lab and I think a guy at auburn has a patent on the process. Ther may be some articles in the literature on this.
Milo
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The most common is etchant nital - 2% nitric acid in methanol. swab the polished surface with the etchant for 2 - 10 seconds. see the metals handbook. the carbide will always be evident, but in addition the ferrite grain boundaries will be evident.
Milo
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I represent Micro Star Diamond Knives, and Technical Manufacturing Corporation and their "MICROg" Vibration Isolation systems. If I can be of any assistance to any one please contact me {tcooper-at-sprintmail.com}
I use a Polaroid SprintScan and am fairly happy with it. Here are a couple of high end scanners that I had gotten info on, but cannot find anymore.
Scitex Smart Scanner: this is a flat bed. Visit Scitex's web site. http://www.scitex.com Ask for info and they will send a demo CD about the scanner. I believe that you are restricted to using a Mac system with this.
Scanview Scanmate 5000 or Scanmate 3000: this is a drum scanner. Visit this site http://www.medgraphix.com/scanmate3000.htm for info on the 3000. It is about $20,000.
I did a search on the web for the above two scanners and found the following sites for vendors of scanners that have a variety of scanners for sale.
"The Scanner Guys, 508-456-9220": http://www.ultranet.com/~bgriffin/ They have a whole bunch of different scanners, but there isn't much info on their page about each one. They want you to call.
Here is another that has new and used scanners of all types including used LeafScan 45's. http://www.promarketinc.com You should be able to find something in your price range with one of these places.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: "edelmare-at-casmail.muohio.edu"-at-sparc5.microscopy.com To: microscopy-at-sparc5.microscopy.com; CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU
Looking for recomendations for a film scanner. The Leaf 45 is no longer in production. So far I have only found the Nikon LS-4500AF and the Polaroid Spirit Scan 45 Pro. Any others? Any recommendations?
Flatbeds are o.k. for 4X5's (we already have one) but would also like to be able to handle 35mm's as well (8x10 } 9x14 flatbed scanners just don't have the resolution for 35mm's).
Thanks again.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
I say make your own! Get the beads (I use Bangs Labs, here in the USA somewhere), dilute the suspension at least 100:1, or even more, drop on slide, let dry, add epoxy and cover glass, let set.....confocal. Used it to evaluate confocals, bought a BioRad ourselves. We are doing volume measurements, so calibrated beads with known std. dev. were useful. We even deformed the beads (heat & pressure) to test ability to find volumes of irregular shapes. Don't mount in oil or water, as the cover glass sticks to lens imersion oil and the little balls scoot all over when moving the stage. Dave Pevear, Houston, Texas
Although this is not a specific microscopy question, somebody of you might have an answer. We just started to study apoptosis in alveolar epithelial cells. We are looking for a reliable method to induce apoptosis in our cell culture system (L132). Has anyone any experience about how to reliably induce apoptosis in the human embryonic lung epithelial cell line L132 ? Any information will be greatly appreciated.
Thank you very much in advance.
Heinz
*********************************************************************** Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
We have a Siemens Elmiskop 102 electron microscope. We use pointed filaments. We distinguish two types of filament : * A one-piece filament where the tip is mechanically thinned * A two-pieces filament where the tip is welded on the loop.
The one-piece filament is the only one which is suitable for our microscope.
Does anyone have any information about this type of filament ?
Thanks in advance for your answers
Sincerelly yours
Philippe Drouillon Solvay Research and Technology Rue de Ransbeek, 310 B-1120 Brussels (Belgium) tel : (0032)2.264.24.47 mailto:philippe.drouillon-at-solvay.com
I am curious to know how does one define Negroid, Caucasian and Mongoloid SCIENTIFICALLY? This smacks of Prof. Rushton's preposterous theories about testosterone and IQ of "races". And statistically, the differences within a defined group is not different ENOUGH to the differences between. So how does one do it? ...by intuition?
Azriel wrote:
My appologies for the lateness in this reply, I have been out of the country. As an old and still practicing classical hair comparison person my views may be seen by many as biased.
The simple answer is that there has been found a relationship between cross sectional shape and racial characteristics. Flat being associated with the Negroid race, oval with the Caucasion race, and round with Mongoloid.
To quote from "MICROSCOPY OF HAIR, A Practical Guide and Manual" put out by the FBI:
"A human hair may be classified according to its racial characteristics as being Caucasian, Negrod or Mongoloid origin. In some instances, the racial characteristics exhibitied by the hair specimine may not be clearly defined indicting the sounce of the particular hair may be of mixed racial origin."
and
"Again, it is pointed out thet even when racial characteristics are not clearly defined, it is significant when these characteristics are consistent between the hair in questiion and the hair of known origin."
Okay, probably more than you wanted to know. Good luck.
Shalom from Jerusalem, Azriel +++++++++++++++++++++++++++++++++++++++++++++++++++++ Major Azriel Gorski, Head Fibers and Polymers Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem, ISRAEL
azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739
What lies behind us and what lies before us are tiny matters compared to what lies within us. Ralph Waldo Emerson ++++++++++++++++++++++++++++++++++++++++++++++++++++
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
You mention that the cylinder of argon is new, it seems possible and likely that this is coincident with the problem and it may be that there is a high water content in the argon. Most labs will have a nitrogen cylinder available and as a check this could be used to see if the problem remains.
Other possibilities are the rotary pump backstreaming oil into the plasma if left to run at ultimate pressure, or an 'O' ring breaking down within the target area.
The operating parameters of 1.4Kv at 50mA would indicate a very old E5000 coater and it is possible that there are vacuum leaks around the target when hot. A good service with 'O' ring replacement may also cure the problem.
Concerning the possibility of non-target material being sputtered through holes in the target, it can occur with some designs of target and target holders. For example another previous Polaron model, the SC500, used a target material bonded to a holding plate using a silver loaded epoxy resin. With this design it was possible to sputter silver and resin components (i.e. contaminantsl) through holes eroded in the target. This is unlikely with the E5000, or with similar sputter coaters that do not use silver bonding material. Although it would seem possible that material from the target holder (i.e.aluminium) could be sputtered through holes in a worn or damaged target, the power (e.g. 1.4Kv at 50mA) and design of such coaters would normally not preclude this.
Best regards M.J.Wombwell Polaron range Business Manager http://www.vgmicrotech.com/polaron-range
I will not get into the anthropological "p'n contest" that has gone on in the recent past on "defining race." As we both know from South Africa and from the experiences of trying to define, scientifically measure, and deal with races in the 1930's/40's and others ... the term and concept of race can be sensitive and has been, to be mild, badly misused.
I will only state that there are accepted and usable definitions, in the common use, if not the scientific. Those definitions have characteristics associated with them.
I feel as a person and even a a scientist, I can be conversant wi, recognize and use those concepts.
Shalom from Jerusalem, Azriel
On Fri, 28 Aug 1998, Quirina Roode wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Azriel Gorski, } } I am curious to know how does one define Negroid, Caucasian and } Mongoloid SCIENTIFICALLY? This smacks of Prof. Rushton's preposterous } theories about testosterone and IQ of "races". And statistically, the } differences within a defined group is not different ENOUGH to the } differences between. So how does one do it? ...by intuition? } } Azriel wrote: } } My appologies for the lateness in this reply, I have been out of the } country. As an old and still practicing classical hair comparison person my } views may be seen by many as biased. } } The simple answer is that there has been found a relationship between cross } sectional shape and racial characteristics. Flat being associated with the } Negroid race, oval with the Caucasion race, and round with Mongoloid. } } To quote from "MICROSCOPY OF HAIR, A Practical Guide and Manual" put out by } the FBI: } } "A human hair may be classified according to its racial characteristics as } being Caucasian, Negrod or Mongoloid origin. In some instances, the racial } characteristics exhibitied by the hair specimine may not be clearly defined } indicting the sounce of the particular hair may be of mixed racial origin." } } and } } "Again, it is pointed out thet even when racial characteristics are not } clearly defined, it is significant when these characteristics are } consistent between the hair in questiion and the hair of known origin." } } Okay, probably more than you wanted to know. Good luck. } } } Shalom from Jerusalem, } Azriel } +++++++++++++++++++++++++++++++++++++++++++++++++++++ } Major Azriel Gorski, Head } Fibers and Polymers Laboratory } Division of Identification and Forensic Science } Israel National Police } Jerusalem, ISRAEL } } azrielg-at-cc.huji.ac.il ICQ User ID No. - 1750739 } } What lies behind us and what lies before us are tiny } matters compared to what lies within us. } Ralph Waldo Emerson } ++++++++++++++++++++++++++++++++++++++++++++++++++++ } } } } \|||/ } (o o) } *----oOO------(_)-OOo---------* } } Quirina I. Roode } PhD student: Cement Chemistry } *-----------------------------* } Department of Civil Engineering } University of the Witwatersrand } P O Wits, 2050, Johannesburg } SOUTH AFRICA } *-----------------------------*Oooo. } ROODE-at-civen.civil.wits.ac.za ( ) } Tel: +27-(0)11-716-2478 (w) ) / } Fax: +27-(0)11-339-1762 (w) (_/ } *.oooO------------------------* } ( ) } \ ( } \_) }
I am going to put my full 2D code for Direct methods (surfaces and TED data) out as Public Domain code soon. I will have a used interface similar to Shelx, and output either of semper images (Fortran files that can be read by other programs if someone writes an interface), "miff" files for ImageMagick or hkl,F,phase.
Any volunteers for testing it? The code is Fortran, pretty much platform independant, but parts of it are specificaly UNIX based.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Philippe Drouillon asked: } We have a Siemens Elmiskop 102 electron microscope. We use pointed } filaments. } We distinguish two types of filament : } * A one-piece filament where the tip is mechanically thinned } * A two-pieces filament where the tip is welded on the loop. } } The one-piece filament is the only one which is suitable for our } microscope. } } Does anyone have any information about this type of filament ?
Energy Beam Sciences offers a proprietary "SG" filament loop, which fits this description. The tip of the filament loop is etched into a "spade" shape. Higher brightness can easily be acheived with a filament of this type.
Please contact me back-channel for additional information.
I have been following this thread with interest. I suspect that by now the sixth grader is wondering how, what he meant as a simple question (I don't think he was thinking of submitting this for publication), has turned into a racially spiked issue about testosterone and IQ. My guess is that by now the student is asking himself a deeper question : "Are microscopists for real ???"
My two cents on the issue.
Jordi Marti
---------- } From: Quirina Roode To: microscopy-at-sparc5.microscopy.com -----------------------------------------------------------------------
Does anyone have any experience with converting an older SEM so that pc-based images can be collected? Two questions: !) What issues are important? 2) Any recommendations on companies that sell the conversions?
Yes, if at all possible, you should have a biohazard hood for dealing with unfixed samples, especially lung samples that may contain such things as TB spores that are easily aerosolized. All other specimens can be moved to a fume hood once they're fixed in glut (except for brain samples which may have spongioform encephalopythy. e.g., Creutzfeld-Jacob disease).
My Surgical Pathology EM Lab processes 300-400 tissue samples per year. All come in glut. They have no biohazard hood. My EM Diagnostic Virology Lab processes around 800 nonfixed specimens per year; all samples go into the biohazard hood where negative stains are processed and tissue samples are placed into glut. Fixed specimens are transferred to the fume hood for further processing.
Sara Miller (Director, Surg Path EM Lab & EM Diag Virol Lab) address below
On Mon, 11 May 1998, Richard Easingwood wrote:
} Date: Mon, 11 May 1998 14:41:57 +1200 } From: Richard Easingwood {richard.easingwood-at-stonebow.otago.ac.nz} } To: MICROSCOPY-at-sparc5.microscopy.com } Subject: Biohazard cabinets for EM labs } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello microscopists, } } We have been given the go ahead to upgrade our electron microscope } preparation laboratory area including the choice of installing new } fumehoods and/or biohazard cabinets. } } At present we have two fume hoods and no biohazard cabinets. We are } thinking of asking for four fume hoods or, alternatively, three fume hoods } and one biohazard cabinet. Our feelings were that the biohazard cabinet } might be safer considering the human biopsy material we deal with, but } maybe it would be no safer than a good fume hood. } } My question is: should we consider a biohazard cabinet in place of a } fumehood given that many of the biohazards dealt with in tha lab are also } in toxic fixatives or solvents? Do other EM labs use biohazard cabinets in } preference to fume hoods? } } (By fume hoods I mean that the fumes are extracted from the room and } released outside whereas a biohazard cabinet filters the air and returns it } to the room). } } Many thanks for any thoughts you might have on this matter. } } Richard } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences } University of Otago } PO Box 913, Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } e-mail: richard.easingwood-at-stonebow.otago.ac.nz } } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I would like to think it would be a simple matter any more. We had a JEOL U3 set up for digital control back in 1981. It was relatively straightforward to find the connections for scan control and to have the scanning hardware set up to work the same voltage range. Unfortunately the computers were not so friendly then, nor were the electronics all that fast. Present D/A cards should be able to do much of that all from within the PC box.
The biggest issues would probably be software, speed and allowance for hysteresis in the scan coils. Taking those in reverse order, I know even our JEOL 840 can be significantly off at high scan speeds, but normal active digital scanning systems will probably not be near that fast. There can be a significant amount of overhead in stepping from point to point so that scans can be slow for 1024 pixels across the image. We find 100 us of dwell per point is plenty from a signal standpoint. I don't know how fast the scanning can reasonably be performed in conjuction with image digitization.
Software is a non-trivial issue. Being a tinkerer, I could probably come up with a hardware solution in a reasonable amount of time. However, the software will probably determine the overall satisfaction with such a system. It is probably the greater investment of energy.
Therefore, IMO, you should probably evaluate commercially available systems to find one with software that meet your criteria (usability, database functions, etc.), and then pursue the question of matching the hardware up to your microscope.
FWIW, we are happily using the Quartz PCI passive imaging system. Rather than taking over control of the beam, the system passively monitors the scan and records the image. I would suppose there is enough latitude in their hardware adjustments to match the raster of virtually any microscope with minimal bother. (Y'all can insert the standard disclaimer here.)
At 12:06 PM 8/28/98 -0500, you wrote: } ------------------------------------------------------------------------ } Does anyone have any experience with converting an older SEM so that } pc-based images can be collected? Two questions: !) What issues are } important? 2) Any recommendations on companies that sell the conversions? } } Thanks, } } Robin Griffin
We have had excellent success with the 4pi System's Spectral Engines for retrofitting old systems as well as new installations. It is a _VERY_ cost competitive option, particularly in terms of flexibility and capabilities. If you already have a scan control interface, hooking up a Spectral Engine is very straightforward. They also have a lot of experience at getting into the depths of older instruments.
http://www.4pi.com 919-489-1757
Bill Heeschen The Dow Chemical Company
} I would like to think it would be a simple matter any more. We had a } JEOL U3 } set up for digital control back in 1981. It was relatively } straightforward } to find the connections for scan control and to have the scanning } hardware } set up to work the same voltage range. Unfortunately the computers } were not } so friendly then, nor were the electronics all that fast. Present D/A } cards } should be able to do much of that all from within the PC box. } } The biggest issues would probably be software, speed and allowance for } hysteresis in the scan coils. Taking those in reverse order, I know } even our } JEOL 840 can be significantly off at high scan speeds, but normal } active } digital scanning systems will probably not be near that fast. There } can be a } significant amount of overhead in stepping from point to point so that } scans } can be slow for 1024 pixels across the image. We find 100 us of dwell } per } point is plenty from a signal standpoint. I don't know how fast the } scanning } can reasonably be performed in conjuction with image digitization. } } Software is a non-trivial issue. Being a tinkerer, I could probably } come up } with a hardware solution in a reasonable amount of time. However, the } software will probably determine the overall satisfaction with such a } system. It is probably the greater investment of energy. } } Therefore, IMO, you should probably evaluate commercially available } systems } to find one with software that meet your criteria (usability, database } functions, etc.), and then pursue the question of matching the } hardware up } to your microscope. } } FWIW, we are happily using the Quartz PCI passive imaging system. } Rather } than taking over control of the beam, the system passively monitors } the scan } and records the image. I would suppose there is enough latitude in } their } hardware adjustments to match the raster of virtually any microscope } with } minimal bother. (Y'all can insert the standard disclaimer here.) } } At 12:06 PM 8/28/98 -0500, you wrote: } } --------------------------------------------------------------------- } --- } } Does anyone have any experience with converting an older SEM so that } } pc-based images can be collected? Two questions: !) What issues } are } } important? 2) Any recommendations on companies that sell the } conversions? } } } } Thanks, } } } } Robin Griffin } }
Cheers, Jordi! Let's get back on topic folks. There may be a place for the discussion that followed posting by the 6th grade student but this list server isn't it.
********************************************************************** Rhonda L. Callender Inorganic & Materials Chemistry Rice University 6100 Main Street MS60 Houston, TX 77005
On Fri, 28 Aug 1998, Marti, Jordi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have been following this thread with interest. I suspect that by } now the sixth grader is wondering how, what he meant as a simple } question (I don't think he was thinking of submitting this for } publication), has turned into a racially spiked issue about } testosterone and IQ. My guess is that by now the student is asking } himself a deeper question : "Are microscopists for real ???" } } My two cents on the issue. } } Jordi Marti } } } ---------- } } From: Quirina Roode } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Shape of Hair Question from a 6th Grade Student } Date: Friday, August 28, 1998 7:03AM } } ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- }
We converted an 1981 ISI DS-130 to a digital acquisition system that we are very pleased with. It has been in use for about 5-6 years. The system (Quartz PCI passive acquisition system) was developed in Canada and is available from:
At 12:06 PM 8/28/98 -0500, rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We converted a 1981 ISI DS-130 to a digital acquisition system that we are very pleased with. It has been in use for about 5-6 years. The system (Quartz PCI passive acquisition system) was developed in Canada and is available from:
At 12:06 PM 8/28/98 -0500, rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If a lab has an SEM or light microscope (up to four), SEMICAPS Image Capture Software will archive your image, paste the image to a report, and use keyword searches for future image retrieval. Depending on your network (internet or intranet) system, SEMICAPS can send the image as a TIF or JPEG file.
SEMICAPS consists of three systems:
The passive system will tap into the video signal of the SEM monitor.
The active system controls the electron beam and is most useful for background noise removal.
The color-capture system will connect up to four light microscopes and includes a "snap feature" that allows each user independent control of the image capture process.
If your image system requires: archival, noise reduction, image processing, measurement, annotation, and printout, SEMICAPS is the system most used.
Give Bruce, Eric or Elie a call if you would like a no-cost demo. 408-986-0121
I am new to the list and to microscopy (just got a used scope for home use). Can anyone help me find an illuminator for an AO student scope of ca. 1950 vintage? This is a black monocular scope with substage condenser. Do I need something particularly for this model or would other illuminators work? If so, how can I tell before ordering? Thanks for your indulgence.
Hi there. Can you email me a pic of the scope? They made several during that time. Please send Pic to couryhouse-at-azonline.com I will check my old catalogs and see if I have something in the Parts box here that might be made to work. thanks Ed Sharpe
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Greetings! You don't say whether your scope (which will give you a lot of enjoyment) has a mirror, or was originally fitted with a substage illuminator. If you have a mirror, you can start with almost anything - even a gooseneck lamp. You can get something more sophisticated when you've developed your own needs and opinions. You do need a book or two. I suggest that you look at the Project MICRO bibliography (address below). There are several good books for adult hobbiests listed; you might start with Nachtigall (Section IIA).
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Discussions at MSA in Atlanta identified a void in high resolution "in vitro" microscopy.
Recent progress made in bio in vitro AFM are meeting this need. Some applications presented clearly point to new microscopy capabilities. For example: 1- time lapsed in vitro imaging of katanin disintegration of microtubule (Vail group UCSF) 2- molecular recognition force microscopy which characterized antibody - antigen binding force in vitro (Schindler group Linz Austria) 3- protein folding force characterization (Trinick group Leeds UK)
Recent innovations in AFM are 1- magnetic AC mode and 2- optimizing the microscope for fluid operation a) under environmental, b) temperature control, and c) electrical properties characterization (elecrophysiology)
These innovations have filled a void in high resolution "in vitro" microscopy.
Please contact me via e-mail or at (800) 819-2519.
This is just a short note to ask your permission to send you details about our latest skywatch gathering for West Central Indiana.
If you are interested and/or would like to attend, reply to this message. We'll send you date, time and location information. If you are not interested, do nothing and you will not receive the information.
Also, if you reply, rest assured that your e-mail address will not be broadcast, sold or otherwise distributed in any manner to any e-mail "SPAM" list. Your reply will return to a real person that will read it and send you the information, if that is what you wish.
I suppose I have to take some of the responsibility of this off- topic racial discussion. Couldn't help it...it is so facinating...and just maybe it is not so off topic, because the issue is whether we can answer these questions under the microscope or can we not. Science isn't all about what we can see and what we can't, it isn't just about data, it's also about interpretation or even inferrence. My apologies...
Cheers, Quirina
\|||/ (o o) *----oOO------(_)-OOo---------*
Quirina I. Roode PhD student: Cement Chemistry *-----------------------------* Department of Civil Engineering University of the Witwatersrand P O Wits, 2050, Johannesburg SOUTH AFRICA *-----------------------------*Oooo. ROODE-at-civen.civil.wits.ac.za ( ) Tel: +27-(0)11-716-2478 (w) ) / Fax: +27-(0)11-339-1762 (w) (_/ *.oooO------------------------* ( ) \ ( \_)
In our research we study particles under an ordinary optical microscope at 20 times magnification. These particles are placed in a so-called micro-reactor that is closed with a glass lid to be able to see the particles. The glass is rather thick (8 mm!) because the reactor is pressurized (max. pressure 30 bar). The problem is that around the particles a 'dispersed cloud' of white light is observed. This light makes it impossible for us to observe the particles in detail.
We are not sure what causes this phenomenon. We think it may have two reasons. First, it may be caused by the diffraction of the light on the glass/air surface, for the glass disturbs the direction of the light. Because of this not all light emitted by a point light source will converge to one point. We didn=92t succeed in deriving the theoretical point spread function. However, we could derive that a point light source was spread out over a large distance, but it also could be derived that most of the light can be focussed to the middle of the =91spot=92 in an infinite intensity top. Second, it may be possible that the glass lid is made of (relatively) low quality glass. It might not be adequately polished, or may contain impurities. We doubt whether this is the reason, however we already ordered a higher quality glass.
Does anybody have another idea what may cause this problem, or even knows it for sure? More important, does anybody know a solution, is it possible to correct for this? Is there anybody else who uses thick cover slips (owing to high/low pressures)? I believe there are lenses that correct for cover slip thicknesses of 1.5 mm, is this because of the same reason? An obvious solution may be a thinner cover slip. Does anybody know an extremely strong material?
Some details about the used microscope: Zeiss AxioTech Vario, 20x LD Epiplan lens (number 442840, N.A =3D 0.4, WD =3D 9.8 mm).
We hope someone can help us with this problem!
Thanks in advance for any help or suggestions you may have.
Ronald Capel
University of Twente Faculty of Chemical Technology PO-Box 217, 7500 AE, Enschede, Netherlands r.capel-at-student.utwente.nl
I have been using the Printerface program, purchased from GW Electronics,6981 Peachtree Industrial Blvd., Norcross, GA 30092-3601. Phone is (800)325-5556, Fax is (770)449-0284. This is a passive system (i.e., it does not take control of microscope operation), and the image can be stored or sent in several file formats. The price was reasonable, and my customers have been pleased with the system.
Leslie
Leslie Eibest Zoology Dept., Box 90325 Duke University Durham, NC 27708 USA (919) 684-2547 leibest-at-duke.edu
I assume you are referring to the ER itself being paracrystalline, and not to crystals of protein or other substances that can occur within the lumen of the ER.
Here are three references that may be of interest (their reference lists will guide you to others):
Anderson RGW, Orci L, Brown MS, Garcia-Segura LM, Goldstein JL, 1983. Ultrastructural analysis of crystalloid endoplasmic reticulum in UT-1 cells and its disappearance in response to cholesterol. J Cell Sci 63:1-20.
Kochevar DT, Anderson ROW, 1987. Purified crystalloid endoplasmic reticulum from UT-1 cells contains multiple proteins in addition to 3-hydroxy-3-methylglutaryl coenzyme A reductase. J Biol Chem 262:10321-10326.
Fringes B, Gorgas K, 1993. Crystalloid smooth endoplasmic reticulum in the quail uropygial gland. Ann Anatomy 175:231-235.
Regards, Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen Department of Anatomy and Cell Biology, Medical Sciences II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 akc-at-umich.edu, Tel (734) 763-1287, Fax (734) 763-1166 http://www-personal.umich.edu/~akc/
} Dear all, we found paracrystalline ER-structures in cells of the } nectaries of plant flowers. Has janyone seen similar structures or knows } about such things documented in the literature? Thanks for help
} Bernward Laube } University of Bielefeld } Faculty of Biology } Department Plant Morphology and Cell Ultrastructure } Universit=E4tsstrasse 25 } Germany 33615 Bielefeld } phone: 0521 1065592 } fax: 0521 1066039 } e-mail: b.laube-at-biologie.uni-bielefeld.de } http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie/abt.1htm#ws } =20
We are looking for sands from around the world to support the GEMS/Proj= ect Micro elementary science program, MICROSCOPIC EXPLORATIONS. If anyone = would like to collect a handful of sand from Cancun for this project it would= be greatly appreciated. If you can send some sand please contact me first= so I can avoid receiving to many samples. The samples can be sent to: =
Dear All: We need to prepare TEM specimen to study a thin film on single crystalline graphite. One of the methods to prepare a sample is to cleave graphite using adhesive tape. Does anybody know the tape which leaves no glue after dissolving? Thanks, Serge.
I did this another way on single crystal MoS2 using low temperature wax and Post-it notes. The MoS2 cleaves like graphite.
I took the MoS2 and waxed it down to a glass microscope slide using low temperature melting wax (glycol phthalate) from SouthBay Technology. After it cooled, I just repeatedly stripped off layers with new areas of Post-it notes until what remained was optically transparent. I dissolved the wax in acetone. I can't remember if I just dipped out the films with a grid or collected them using a loop. I think that I tried both. You could use a variation if you lowered the acetone bath level so that the films were collected on a grid support plate.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Serge Oktyabrsky To: Microscopy-at-sparc5.microscopy.com
Good day all,
Here's a question for the TEM/STEM crowd (in my case a Philips CM30).
I'm looking for advice on what is apparently an alignment problem that results in the shift of my PEELS signal, up and down on the energy scale (by about 1.5-2 eV) as the STEM beam is rastered across a line. Up 'till now, I thought that I had figured out how to do a decent STEM alignment (decent resolution in STEM images up to about 400kX) but apparently something is still off. I have tried numerous realignments of the pivot points and rotation centering- along with the DESCAN set up- with little luck. Does anyone have any advice before I have to start thinking about what I'm doing?
Specifics; Philips CM30 (300kV) Gatan 666 spectrometer. Following STEM alignment procedures given from Philips (if anyone wants details, I can write them out and I'd be happy to discuss them since the problem could be a result of interpretation). Using DESCAN mode as described in CM30 instruction manual for STEM-PEELS. -checking the voltage rotation centering and beam pivot points... is there a problem going from doing this on the view screen which is at a different height from the spectrometer entrance aperture?
I've been at this for a day now, and while I still think I can figure it out, I'd be extremely happy to get some advice if it is easy of obvious to anyone with more STEM-PEELS experience. I'd be happy to discuss via email, or if anyone wants to talk me through it, I'd be happy to return a page.
Brendan Foran, Ph.D. Materials Analysis Group SEMATECH Austin, TX
phone: 512-356-3936 (never there, but messages can be left on voicemail) pager: 1-888-731-4459 ________________________________________________________________________ ___ For fun: ...I can only be glad that I do not use the "CM30" described at: http://www.neatworld.com/incont.htm ..parallel universes?
[skip] } An obvious solution may be a thinner cover slip. Does anybody } know an extremely strong material? } Either fused quartz or, better yet, saphire are extremely strong. I don't know, however, what area one can use to contain the pressures you're working with. Good luck. Yours, Bill Tivol
With 120 film (2 and 1/4 inch square), what does the 120 refer to? It can't be 120 mm frame size. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
A parallel plate of glass will introduce spherical aberration into a diverging beam. This is why high NA lenses are designed with a cover slip in mind if one is to be used. In your extreme case you will have roughly 200 microns of spherical aberration if your particles are in air, about half that if they are in liquid.
This is a tough problem. One solution would be to make a window from a concentric meniscus lens--where both surfaces have the same center of curvature. Then if the particles are at the center of curvature and the rays hit the window perpendicular to its surface you should eliminate the spherical aberration, or at least minimize it. This will make a much stronger window, although for your geometry it will be small.
If you have liquid in your chamber a plano-convex lens might suffice.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
My guess would be the image is suffering from sherical aberation. Perhaps you could find an objective off an inverted scope that has a correction collar that will allow you to adjust the ojective to the cover glass thickness. I doubt it can adjust to 8mm however.
bob Derm Imaging Center U of W
On Mon, 31 Aug 1998, Ronald Capel wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Dear all, } =20 } In our research we study particles under an ordinary optical } microscope at 20 times magnification. These particles are } placed in a so-called micro-reactor that is closed with a } glass lid to be able to see the particles. The glass is rather } thick (8 mm!) because the reactor is pressurized (max. } pressure 30 bar). The problem is that around the particles a } 'dispersed cloud' of white light is observed. This light makes } it impossible for us to observe the particles in detail. } =20 } We are not sure what causes this phenomenon. We think it may } have two reasons. } First, it may be caused by the diffraction of the light } on the glass/air surface, for the glass disturbs the direction } of the light. Because of this not all light emitted by a point } light source will converge to one point. We didn=92t succeed in } deriving the theoretical point spread function. However, we } could derive that a point light source was spread out over a } large distance, but it also could be derived that most of the } light can be focussed to the middle of the =91spot=92 in an } infinite intensity top. } Second, it may be possible that the glass lid is made of } (relatively) low quality glass. It might not be adequately } polished, or may contain impurities. We doubt whether this is } the reason, however we already ordered a higher quality glass. } =20 } Does anybody have another idea what may cause this problem, or } even knows it for sure? More important, does anybody know a } solution, is it possible to correct for this? Is there anybody } else who uses thick cover slips (owing to high/low pressures)? } I believe there are lenses that correct for cover slip } thicknesses of 1.5 mm, is this because of the same reason? An } obvious solution may be a thinner cover slip. Does anybody } know an extremely strong material? } =20 } Some details about the used microscope: Zeiss AxioTech Vario, } 20x LD Epiplan lens (number 442840, N.A =3D 0.4, WD =3D 9.8 mm). } =20 } We hope someone can help us with this problem! } =20 } Thanks in advance for any help or suggestions you may have. } =20 } Ronald Capel } =20 } University of Twente } Faculty of Chemical Technology } PO-Box 217, 7500 AE, Enschede, Netherlands } r.capel-at-student.utwente.nl } =20 } =20 } =20
In a message dated 98-08-31 17:50:29 EDT, bozzola-at-siu.edu writes:
{ { With 120 film (2 and 1/4 inch square), what does the 120 refer to? It can't be 120 mm frame size. Thanks.
This interesting little bit of confusion dates back to the commercialization of cameras and films, and we have Kodak to thank.
In the beginning (about 1898-1910) not all films fit all cameras, and the situation became worse as new cameras were introduced. Kodak decided to start numbering all of their films which were wound on flanged spools, beginning with the number 101, which was a standard spool of film that fit a specific camera. We still use this convention today. Several of the intermediate types have been dropped, and I think 127 film is scheduled to bite the dust in the year 2000.
You can learn more than you want about this bit of history by visiting:
Hi Folks, I have been trying to use EMbed-812 for TEM on C.elegans. I am using an ultracut E, with a glass knife and I just can not seem to get any thin sections using this resin.Poly/Bed 812 has worked fine, but is a little too viscous. I have tried both medium and hard mixtures to no avail. Does anyone have any ideas, I don't have access to a diamond knife.
THANK-YOU to all those who responded to my query on where to purchase slides with fluorescent beads on them for use in confocal calibration. I was asked by a few people to let others on the list know where to purchase these slides from. I received responses from many people telling us how to make the slides ourselves but only two people responded with actual suppliers of these slides. They were as follows;
1. "Molecular Probes, Inc. sells their "MultiSpeck Kit" Catalog # M-7900, consisting of three slides, each wiht two zones. List price is US$125." (thanks to Micheal Nesson). 2. "Polysciences make fluorescent beads already on slides Cat. # 24016 Confocal Microscopy-Multifluorescent Adjustment and Calibration Kit." (thanks to Jane Woodruff).
I hope this is of use to other people out there,
Sarah Ellis
Trescowthick Research Centre Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett Street Melbourne 8006 Victoria Australia
At 16:37 31/08/98 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
width max picture width code
16 mm 13mm 110 35mm 24mm 135 35mm 28mm 126 single perforation 46mm 40mm 127 62mm 60mm 120 OR 620 OR 220 (double length)
***************************************************** Mel Dickson, Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia