} I have found some references to books on TEM history by Marton (1968) } and Hawkes (1985) but don't have these on hand.
I have a book that was published at the time of the International Congress on EM in Kyoto, Japan, in 1986, entitled "History of Electron Microscopes 1986" edited by Hiroshi Fujita. This gives a lot of information on the history of EM, mainly from a Japanese perspective. I haven't seen whether or not this has the answers to your specific questions but if there is someone near you who has a copy of this book you will find much interesting information in it. Please let me know if you cannot get hold of a copy.
Robin H Cross (Mr) Director : Electron Microscopy Unit Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 email: r.cross-at-ru.ac.za http://www.ru.ac.za/emu/index.htm
** remember ICEM-15 in Durban in September 2002 **
I am not a forensic scientist so my suggestions are general - therefore I am uncertain how the methods I give could be made robust enough for use in court. I assume that IPA is isopropyl alcohol?
Starch should be positive with the periodic acid Schiff method (PAS) and may be 'confirmed' as such by using an amylase control. Starch should also show 'Maltese Cross' birefringence under polarised light. The cellulose should also be PAS positive, resist amylase and show 'linear' birefringence.
The protein is a little more difficult to confirm absolutely but if these are skin cells they will contain cytokeratins - thus giving the possibility of using immunocytochemistry with a pancytokeratin antibody.
Good luck!
PS Why is an IR/Raman engineer doing forensic work on paper/cells?
At 16:07 2001-02-28 -0800, HAMMOND,LOMA (HP-Corvallis,ex1) wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
Congo red will stain cellulose red, and can also be viewed by fluorescence using a green (RITC-type) excitation filter. Cellulose and starch are also birefringent, which you could demonstrate by using crossed polarizers. A 1/4 wave or 1-wave plate may make this easier to see. The fluorescence of Cellulose stained with congo red is also polarized because the CR molecules line up on the oriented cellulose microfibrils. As has already been mentioned starch and cellulose stain with periodate-Schiff. If you have access to a fluorescence microsocope you might also try a fluorescence variant of PAS, by replacing schiff reagent with Lucifer Yellow CH. View with FITC-type excitation.
Iodine/potassium iodide will stain starch blue/black.
Chris
----- Original Message ----- } From: "HAMMOND,LOMA (HP-Corvallis,ex1)" {loma_hammond-at-hp.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 01, 2001 12:07 AM
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Fungal people, Although vapor fixation works sometimes and immersion fixation also works sometimes, often fungi samples are too delicate to handle with routine techniques. Just immersion in an aqueous solution can damage the structures. Repeated washings also can wash away delicate conidia. CPD causes shrinkage which may not be a major concern with some samples but can be a real problem with delicate ones like fungal hyphae. Really the very best way to deal with many fungi samples is cryo SEM. True you may have to coat a bit longer than normal, etc. but the structures, when instantly frozen, are most likely to be similar to the living state. Unfortunately many people do not have access to Cryo -SEM equipment so must settle for the next best approach....fixation and drying by one of the methods already mentioned.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
Microlisters,
While the simpler and less time consuming suggestions already mentioned in this thread may work in some cases, I found out a long time ago that the only way to deal with the "fungal jungle", eg. fungi cultured on agar plates, is to hit 'em with the ol' OTO method, preceded by glutaraldehye fixation. Quick outline is as follows:
Glut. fixation, your favorite buffer Buffer rinses Osmium tetroxide, 2% Water rinses Thiocarbohydrazide, sat. soln.,filtered water rinses Osmium tetroxide, 2% Water rinses dehydration series, EtOH, or acetone CPD metal coating? Maybe, maybe not. Try without first.
This eliminates the charging, and fixation is quite good.
Classic reference for this method:
"Ligand-mediated osmium binding:Its application in coating biological specimens for scanning electron microscopy". Kelley, Robert O., A.F. Dekker and John G. Bluemink. J. Ultrastrucute Research 45:254-258 (1973).
See also:
"Non-coating techniques to render biological specimens conductive/1980 update". Judith A. Murphy. Scanning Electron Microscopy/1980 Vol.I, p. 209-220.
You might try exposing your non-aquatic sample (bread molds, mushrooms, etc. ) to vapors of Osmium tetroxide for several hours ( a few drops of 4% OsO4 on the lid of a very small container with your sample inside or the lid over the sample directly), then air dry, mount, coat and view. As with any sample, the drying artifacts vary with the fungus. Some things look great.
good luck
Steve } ----------------------------------------------------------------- } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks...
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
As others have pointed out, phase halo is easily demonstrated. Shading-off is a different matter, at least for me. Both artifacts are the result of incomplete segregation of the direct and diffracted light to the conjugate and complementary areas respectively of the phase plate. Shading-off is described in "Advanced Light Microscopy" vol. 2 pg. 15-20 by Maksymilian Pluta. Shading-off appears as a brighter central area becoming darker toward the edge in positive phase contrast, and just the reverse for negative phase contrast. Pluta has illustrations of this on pages 18 and 19, but he doesn't say what the object is. It looks like a crystal of some kind.
My point in bringing this up is to solicit others ideas on how best to teach students (as well as ourselves) to interpret the phase contrast image. The phase image has these artifacts superimposed on it. The phase halo can mislead a novice but actually help an expert. Shading-off is more subtle in a complex specimen than it is in Pluta's example, and I have never seriously considered how it affects the phase image. I wonder if there is a biological sample that clearly shows both artifacts? Any thoughts and suggestions on this?
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 http://www.pathology.med.unc.edu/path/microscopy/welcome.htm
Email: kuznetsv-at-geo.komisc.ru Name: Chuprov Georgy School: The Institute of Geology State: Republic of Komi
Question: Please, tell me. Where I can find a computer program for plot of stereografic projections of optical axes (were tested by optical microscop)?
I have one of those very basic "thought" questions on a procedure that I've always taken for granted, but am now not so sure about. I had been trained from Day One to always focus a TEM with the beam at crossover, then to spread the beam until the exposure intensity is reached. The idea seems to be that small focusing errors at crossover will be corrected as the beam is spread and "depth of focus" is increased somehow.
The reason I'm asking is that in recent years, almost no one else I've run across focuses in this way, preferring instead to focus with the beam already spread to the point at which the photograph will be taken (at least as long as the image is bright enough to see for focusing). Out of curiosity, I ran tests using both methods and have noted no clear differences.
Does anyone have any thoughts on this? Any ideas or theories about why one method would be superior to the other?
Sincerely, Curious in Columbia
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Dr. Ahlstrand's technique is excellent for imaging hyphal structures and some spore bodies while the stick on method of Dr. Mary Mager works well for certain individual spores. However, imaging of fruiting body structures (conidiophores, etc.) are often best prepared by using Osmium vapor fixation followed simply by air-drying (AD). One can also add minimal solvent exchange before AD, but some structural collapse will be observed in any event. "One size does not fit all" for fungi, sometimes even within one species.
We have a new ESEM now and hopefully combinations of low vacuum technology and various wet modes will improve imaging for our mycologists.
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Microlisters,
While the simpler and less time consuming suggestions already mentioned in this thread may work in some cases, I found out a long time ago that the only way to deal with the "fungal jungle", eg. fungi cultured on agar plates, is to hit 'em with the ol' OTO method, preceded by glutaraldehye fixation. Quick outline is as follows:
Glut. fixation, your favorite buffer Buffer rinses Osmium tetroxide, 2% Water rinses Thiocarbohydrazide, sat. soln.,filtered water rinses Osmium tetroxide, 2% Water rinses dehydration series, EtOH, or acetone CPD metal coating? Maybe, maybe not. Try without first.
This eliminates the charging, and fixation is quite good.
Classic reference for this method:
"Ligand-mediated osmium binding:Its application in coating biological specimens for scanning electron microscopy". Kelley, Robert O., A.F. Dekker and John G. Bluemink. J. Ultrastructure Research 45:254-258 (1973).
See also:
"Non-coating techniques to render biological specimens conductive/1980 update". Judith A. Murphy. Scanning Electron Microscopy/1980 Vol.I, p. 209-220.
You might try exposing your non-aquatic sample (bread molds, mushrooms, etc. ) to vapors of Osmium tetroxide for several hours ( a few drops of 4% OsO4 on the lid of a very small container with your sample inside or the lid over the sample directly), then air dry, mount, coat and view. As with any sample, the drying artifacts vary with the fungus. Some things look great.
good luck
Steve -----------------------------------------------------------------
Dear rad0, I have looked at fungus and fungus spores on my SEM and they are very simple to prepare. Just stick them to a stub by sticky tab or glue and then gold coat. They are very robust and they don't need any fixing or dehydration.
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 e-mail: mager-at-interchg.ubc.ca
} ----------------------------------------------------------------- } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks...
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
You might guess from the lack of response that quantitative stereoscopy using TEM is not that popular a pastime. I was involved some 20 years ago and can offer some comments which were relevant then.
There is a big difference between TEM and SEM stereo analysis. In SEM images you have a lower surface that can be seen easily, in TEM images you have a view through both surfaces and the specimen, the surfaces are very hard to distinguish. It is relatively easy to calculate the height from a fixed surface as in a SEM micrograph. Calculating the height from an 'invisible' surface, as in a TEM micrograph, is very difficult. It's OK to think of something like a dislocation running through the specimen and then assume that it ends at either surface but very few examples go through from surface to surface and thus are considerably more difficult.
Height measurements are made from stereo pairs by measuring the parallax between the two tilted images. Provided the tilt axis is known and the magnifications corrected, for any change in focus current, then the height difference between two points can be calculated. This is a long way from reconstructing a 3D image.
It is relatively easy to view stereo images using a stereo viewer but not everyone can do it. You need two eyes of equal strength and a certain type of brain. From memory the mapmakers said 50% of the population could see stereo properly and 10% could make accurate measurements.
There were programs that would allow 2 stereo micrographs to be placed side by side on a graphics tablet and then the same points marked on both micrographs. The height differences between these points were then calculated using the first pair of points as a reference height. I don't know if these are still available for modern computers (they ran on an Apple 11+).
Stereo viewers are available that will project a 'floating' point of light in the specimen that you can position onto a feature and then read out the height from a micrometer (it will need to be scaled to get the real height). We modified one of these viewers to record and calculate these heights (and X,Y coordinates) automatically), but it it still a lot of work to produce even a simple 3D image.
I am not aware of any image recognition program that will analyse your two images and produce a 3D image from them. (Well if that doesn't get details sent in nothing will!)
If you want some references then in the mid 70s Hudson and Makin descibed the how to select the tilt angle taking into account the film thickness and the final magnification. Much of the work at that time was being done by Alan Boyde. I may be more use with references when I get my office back - contact me off list in a few days if you want 20-25 year old refences.
Good luck, Ron
On Wed, 28 Feb 2001 10:03:56 -0700 Rodney McCabe {rmccabe-at-lanl.gov} 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: } } I am interested in doing quantitative stereo microscopy using a TEM and am } looking for software that may be helpful in such an endeavor. I anticipate } the primary application will be examining the spatial arrangement of linear } defects. I have not been able to find a software designed directly for } quantitative stereo TEM. I have come across one commercial software that } can be used for SEM surface topography that may be applicable. It seems } like there must be other software packages out there used for applications } such as surface mapping, electron tomography, or even astronomy that could } be useful. Also, as I am new to stereo microscopy, any words of wisdom } would be appreciated. } } Thanks, } } Rod } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
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Pete Augustus Caswell Technology
Hi Listers,
I'm preparing a talk which is to include a timeline giving dates and names for some relevant TEM developments. I'm trying to find answers to two trivia questions with great importance to microscopy:
1) Who was the first to knowingly image a dislocation in a TEM, when was it done and where was it published? On this, I have found references as far back as an article by Heidenreich in J. Appl. Phys, 1949, but don't have this journal going back that far.
2) Who developed the solid state detector for energy analysis of x-rays (EDS), when and where published?
I have found some references to books on TEM history by Marton (1968) and Hawkes (1985) but don't have these on hand.
Thanks,
Wharton
*********************** Wharton Sinkler UOP LLC Des Plaines, IL
For fragile conidiophores and their associated conidial chains, it is still possible to use critical point drying after vapor fixation with OsO4.
A fairly simple dehydration technique that avoids the solution changes -- and turbulence -- that often lead to loss of the conidia from aerial conidiophores (and the air drying that often does the same), is described at Can. J. Microbiol. 29:653-658 (1983).
It's an adaptation of a slick idea published in Naturwissenshaften 17:402-403, for TEM, in 1962(!).
I always "heard" to focus the image with the beam in an overfocus condition (past crossover) for improved beam coherence. I find it difficult to focus at crossover anyway. Also, I don't think you can accurately assess focus or astigmatism with the beam at intensities suitable for most photography. Depends on the magnification and a host of other things, of course, such as your exposure time. I don't think there's any justification to focus at exactly the same intensity at which you will photograph. I too await further enlightenment from the list....
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. [SMTP:TindallR-at-missouri.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 } -----------------------------------------------------------------------. } } } Hi all, } } I have one of those very basic "thought" questions on a procedure that I've } always taken for granted, but am now not so sure about. I had been trained } from Day One to always focus a TEM with the beam at crossover, then to } spread the beam until the exposure intensity is reached. The idea seems to } be that small focusing errors at crossover will be corrected as the beam is } spread and "depth of focus" is increased somehow. } } The reason I'm asking is that in recent years, almost no one else I've run } across focuses in this way, preferring instead to focus with the beam } already spread to the point at which the photograph will be taken (at least } as long as the image is bright enough to see for focusing). Out of } curiosity, I ran tests using both methods and have noted no clear } differences. } } Does anyone have any thoughts on this? Any ideas or theories about why one } method would be superior to the other? } } Sincerely, } Curious in Columbia } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } }
University of Central Florida Advanced Materials Processing and Analysis Center (AMPAC) Materials Characterization Facility (MCF)
The Advanced Materials Processing and Analysis Center (AMPAC) is seeking candidates for up to two positions as Assistant in Research to support the Materials Characterization Facility (MCF). Each individual must have at least a Bachelor degree from an accredited institution in an appropriate area of study related to surface science, materials characterization, ion beam analysis, or vacuum science, and preferably three years of experience in any of the above mentioned areas. A graduate degree in the aforementioned areas is preferred. The MCF houses an RBS, a heavy ion beam system, an Auger, an XPS, two FIBs and four SIMS instruments. Opportunities also exist with collaborative interaction with Lucent Technologies, Orlando, which houses an Auger, four FIB systems including a FIB/SIMS and two dedicated SIMS tools.
The person will be responsible for establishing, maintaining and operating ion beam and/or surface science laboratory infrastructure. Depending on the experience, the person should be able to instruct students, faculty, and staff on the use of equipment and in the interpretation of data, conduct independent research and develop surface analysis techniques. AMPAC is particularly interested in individuals desiring an academic setting to further their professional goals. Salary will be commensurate with experience. The applications will be reviewed beginning March 19, 2001 and will continue to be reviewed until the position is filled.
Applicants should send a vitae and a list of three references to Dr. Lucille Giannuzzi, Director MCF, 12443 Research Parkway, Suite 305, Orlando, FL 32826. UCF is an equal opportunity/affirmative action employer. As an agency of the State of Florida, all application materials and selection procedures are available for public review.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
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I have focused at a higher beam intensity than I use for exposing the micrograph for many years, and, in fact continue to do so on our Philips EM201. When we got a new Philips TwinLens CM12S in December of 1988, we started to have problems with slightly out of focus micrographs across the board - all experienced biological microscopists - about 10-12 of us - had the same problem. Philips, to their credit, tried repeatedly to find and solve the problem. They sent high level engineers, applications experts, attached a high mag TV camera, changed all of the high tension cabling, insulator, wehnelt, column liner, etc. over the course of nearly a year. Towards the end of 1989, in desperation and because the biological TEM applications experts were all busy, a materials science applications engineer came to the lab and made beautiful, perfectly focused micrographs. We could not duplicate his performance. Finally, he asked to watch one of our most experienced and well published microscopists work. When the Professor increased beam intensity to focus and stigmate, the applications engineer went ballistic! he maintained that the focus MUST be done at the intensity at which the micrograph is made. We did argue, but also agreed to test the hypothesis. He was, of course, correct (IN THIS PARTICULAR CASE). As we later found out, changing the C2 lens current also has a subtle effect on the objective lens current, changing focus enough to cause problems. Our service engineer was later able to demonstrate the effect (although it doesn't show up when monitoring the lens current page) and this strange phenomenon was written off to a design compromise which, apparently, physicists and materials scientists are used to and expect, but which biologists and cell biologists had never heard of. Something to do with short focal length objective lenses, low contrast which makes focusing more difficult, and compromises required to allow STEM and TEM to co-exist in the same column and be switched back and forth with a minimum of re-alignment. Anyone else have this experience?? William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
I spread the beam as much as possible in order to be able to see the changes during the focusing (usually the intensity is less than necessary to make a photograph, so, I have to condense the beam a little bit for photographing). I am using "three-chick" technique: rotating focus-control knob in both directions to find the position when you will clearly see the transition: "owerfocus-infocus-underfocus" on the background's granules. This technique works for me from x40K and up. At the lower magnification, I am using wobbler to focus (there is a "blind spot" at x30-40K). The "three-chick" technique is a good test how your instrument is aligned. The number of "clicks" depends from current setup and model of coarse. I simply could not focus correctly if the beam is condensed too much.
Sergey
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Well, I got my answer----right from one of the people who initially trained me back in the '80's (and, in fact, is responsible for me being in the field in the first place----that'll teach him).
Here is John Bozzola's reply to my post on focusing TEM's at beam crossover versus a defocused beam: "(T)his is a holdover from (our Hitachi) HU11A when illumination was a problem at the higher mags and when no focus wobbler was available for low mag work. On the other hand, it still DOES work for individuals who have diminishing eyesight and need the extra illumination and minimal depth of field. Unfortunately, it greatly increases the possibility of specimen damage and should be used only on hardy specimens.
"The bottom line is that if you have normal (or acute) vision, you won't need to do this in modern TEMs. Of course, I sometimes still "check" my focus by doing this..... old habit, I guess."
Now this is very interesting to me, because the first TEM I ever used was the Hitachi HU11AB. I was trained to focus carefully at crossover, then defocus until the proper exposure intensity was reached. Somehow over the years, I ended up with the notion that there was a property of electron "optics" that made this the preferred method of focusing because it minimized focusing errors by increasing the depth of focus. I really don't recall where this notion came from, but I trained dozens of students and others in this way of focusing. Now I wonder how many others were trained by them to do the same thing and will be just as stubborn about it as I have been.
Probably not much harm done, really, but this reminds me of one of my favorite stories about a woman cooking a ham for a family reunion picnic. Her daughter watched her mother cut off the end of the ham before putting it in a pan in the oven, and she asked why she did that. Her mother responded that that's how she was taught by HER mother, but she didn't really know why. She became curious, and at the reunion she asked her mother why she had always cut the ends off hams before cooking them. The response was that she had also been taught that way by her mother. Both curious now, they hunted up Great-grandmother at the picnic and asked her why she had cooked hams with the ends cut off. Great-grandmother answered, "I only had one pan and it was too short for the ham."
Lesson learned. Again.
Cheers, Randy
Randy Tindall Electron Microscopy Core University of Missouri
In terms of contrast transfer theory (i.e. when linear imaging allows approximating the imaging process as convolution with a point spread function), the lens transfer functon doesn't depend on convergence. So for example you shouldn't effect the defocus of the image by slight changes of the illumination convergence.
However, the convergence imposes a damping on the total contrast transfer, which gets more severe when convergence is greater. For details on this 'spatial coherence envelope' see any text on HREM. Because of this, the image quality will improve if you record with a less condensed beam (better coherence).
With a field emission gun, you won't be able to image at crossover, because the source will almost certainly be too small - these guns give the most coherent illumination available.
For a thermionic gun, the limiting factor will be: if the convergence is too great at crossover, you won't see much because everything will be blurred out by the coherence limitation of the incident irradiation. If this is the case, you should either change condenser apertures to decrease convergence (then you can focus, and/or stigmate at crossover), or spread the beam to improve coherence.
As far as I can see, there is no reason why one should or should not adjust or even record images at crossover (except that if the filament is a bit undersaturated, you will get uneven illumination). Getting the best possible coherence is important, but it doesn't really matter how one achieves this optically. Decreasing intensity is therefore good, to the extent that sample drift doesn't become too much a limiting factor.
Overfocusing the illumination rather than underfocusing is better, I would say (based on experience). I'm not sure why - perhaps some of the energy spread gets filtered out more effectively - at any rate for the same size beam one has more intensity underfocused than overfocused, so something is being cut out by overfocusing.
Wharton
} -----Original Message----- } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } Sent: Thursday, March 01, 2001 2:30 PM } To: 'Microscopy-at-MSA.Microscopy.com' } Subject: RE: Focusing } } ------------------------------------------------------------------------ } 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 always "heard" to focus the image with the beam in an overfocus } condition } (past crossover) for improved beam coherence. I find it difficult to } focus at } crossover anyway. Also, I don't think you can accurately assess focus or } astigmatism with the beam at intensities suitable for most photography. } Depends on the magnification and a host of other things, of course, such } as } your exposure time. I don't think there's any justification to focus at } exactly the same intensity at which you will photograph. I too await } further } enlightenment from the list.... } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } [SMTP:TindallR-at-missouri.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 } } -----------------------------------------------------------------------. } } } } } } Hi all, } } } } I have one of those very basic "thought" questions on a procedure that } I've } } always taken for granted, but am now not so sure about. I had been } trained } } from Day One to always focus a TEM with the beam at crossover, then to } } spread the beam until the exposure intensity is reached. The idea seems } to } } be that small focusing errors at crossover will be corrected as the beam } is } } spread and "depth of focus" is increased somehow. } } } } The reason I'm asking is that in recent years, almost no one else I've } run } } across focuses in this way, preferring instead to focus with the beam } } already spread to the point at which the photograph will be taken (at } least } } as long as the image is bright enough to see for focusing). Out of } } curiosity, I ran tests using both methods and have noted no clear } } differences. } } } } Does anyone have any thoughts on this? Any ideas or theories about why } one } } method would be superior to the other? } } } } Sincerely, } } Curious in Columbia } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } }
We're analysing Cu wire here for purposes better known to ourselves. We are noticing, but not able to image interesting variations in the concentration of impurities, which suggests to us, either that the original material was inhomogeneous, or that drawing the wire has caused some kind of exsolution process. Have any of you encountered smilar things in metals? Refs? Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
To add to Wharton's explanation the reason overfocus is better than underfocus is that he has a strong objective lens (as most are these days). Many years ago underfocus would have been better.
What we are trying to achieve is parallel illumination not convergent (or divergent). Modern TEMs have strong objective lenses and use the prefield (field above the specimen) as part of their probe forming optics thus if there is a crossover above the specimen (overfocus)the prefield converges the diverging rays from this to form a parallel probe. In underfocus conditions the crossover is below the specimen and the probe on the specimen is made more convergent by the objective lens prefield.
If you have a Lorentz (low field for magnetic work) pole piece or an old instrument (pre 1970ish) the objective lens is weak and does not have this prefield effect so underfocus is more parallel.
Regards, Ron
On Thu, 1 Mar 2001 15:30:22 -0600 "Sinkler, Wharton" {WSinkler-at-uop.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 } -----------------------------------------------------------------------. } } } } In terms of contrast transfer theory (i.e. when linear imaging allows } approximating the imaging process as convolution with a point spread } function), the lens transfer functon doesn't depend on convergence. So for } example you shouldn't effect the defocus of the image by slight changes of } the illumination convergence. } } However, the convergence imposes a damping on the total contrast transfer, } which gets more severe when convergence is greater. For details on this } 'spatial coherence envelope' see any text on HREM. Because of this, the } image quality will improve if you record with a less condensed beam (better } coherence). } } With a field emission gun, you won't be able to image at crossover, because } the source will almost certainly be too small - these guns give the most } coherent illumination available. } } For a thermionic gun, the limiting factor will be: if the convergence is too } great at crossover, you won't see much because everything will be blurred } out by the coherence limitation of the incident irradiation. If this is the } case, you should either change condenser apertures to decrease convergence } (then you can focus, and/or stigmate at crossover), or spread the beam to } improve coherence. } } As far as I can see, there is no reason why one should or should not adjust } or even record images at crossover (except that if the filament is a bit } undersaturated, you will get uneven illumination). Getting the best } possible coherence is important, but it doesn't really matter how one } achieves this optically. Decreasing intensity is therefore good, to the } extent that sample drift doesn't become too much a limiting factor. } } Overfocusing the illumination rather than underfocusing is better, I would } say (based on experience). I'm not sure why - perhaps some of the energy } spread gets filtered out more effectively - at any rate for the same size } beam one has more intensity underfocused than overfocused, so something is } being cut out by overfocusing. } } Wharton } } } } -----Original Message----- } } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } } Sent: Thursday, March 01, 2001 2:30 PM } } To: 'Microscopy-at-MSA.Microscopy.com' } } Subject: RE: Focusing } } } } ------------------------------------------------------------------------ } } 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 always "heard" to focus the image with the beam in an overfocus } } condition } } (past crossover) for improved beam coherence. I find it difficult to } } focus at } } crossover anyway. Also, I don't think you can accurately assess focus or } } astigmatism with the beam at intensities suitable for most photography. } } Depends on the magnification and a host of other things, of course, such } } as } } your exposure time. I don't think there's any justification to focus at } } exactly the same intensity at which you will photograph. I too await } } further } } enlightenment from the list.... } } } } Matt } } } } Matthew J. Lynn } } Center for Advanced Microscopy } } University of Miami } } (305)284-4736 } } mlynn-at-miami.edu } } } } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } } [SMTP:TindallR-at-missouri.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 } } } -----------------------------------------------------------------------. } } } } } } } } } Hi all, } } } } } } I have one of those very basic "thought" questions on a procedure that } } I've } } } always taken for granted, but am now not so sure about. I had been } } trained } } } from Day One to always focus a TEM with the beam at crossover, then to } } } spread the beam until the exposure intensity is reached. The idea seems } } to } } } be that small focusing errors at crossover will be corrected as the beam } } is } } } spread and "depth of focus" is increased somehow. } } } } } } The reason I'm asking is that in recent years, almost no one else I've } } run } } } across focuses in this way, preferring instead to focus with the beam } } } already spread to the point at which the photograph will be taken (at } } least } } } as long as the image is bright enough to see for focusing). Out of } } } curiosity, I ran tests using both methods and have noted no clear } } } differences. } } } } } } Does anyone have any thoughts on this? Any ideas or theories about why } } one } } } method would be superior to the other? } } } } } } Sincerely, } } } Curious in Columbia } } } } } } Randy Tindall } } } EM Specialist } } } Electron Microscopy Core Facility } } } W122 Veterinary Medicine } } } University of Missouri } } } Columbia, MO 65211 } } } Tel: (573) 882-8304 } } } Fax: (573) 884-5414 } } } Email: tindallr-at-missouri.edu } } } Web: http://www.biotech.missouri.edu/emc/ } } } } } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
we are looking for a working Nanoscope III (III or IIIa) controller together with the PC boards (Digital Instruments, Santa Barbara, USA). If somebody is going to clean out its lab please contact us directly by phone, fax or e.mail
With best regards,
Dr. Dmitry Cherny, PhD, Dr.Sc.
MPI for Biophysical Chemistry, Dept. of Molecular Biology am Fassberg 11, D-37077 Gottingen, Germany tel: +49(0) 551 201 1383; fax: +49(0) 551 201 1467 e.mail: dtcherny-at-mpc186.mpibpc.gwdg.de
does anyone have any suggestions on how to assess hydronephrosis with paraffin sections of kidney using a quantitative technique. equipment we currently use for other tissues include microscope, spot camera, digitizing tablet, camera lucida, SigmaScan, and image processing software plugins for adobe photoshop,
Nonuniform distribution of impurities in metal wires is not uncommon. There may be a core-to-surface variation, which may be caused by contamination (or de-contamination) of the surface layer during drawing or annealing . Variations in impurity concentration may also relate to original inhomogeneity in the billet prior to drawing. Segregation of impurities to dislocation-rich deformed areas is also a possibility.
What are the impurity elements that you are seeing? How are the concentration variations distributed spatially? (Radial gradients, irregular patches smaller than wire diameter or larger than wire diameter, wire bends vs. straight segments, or what?)
-------Roy ==================================== Roy Arrowood, Associate Professor Metallurgical and Materials Engineering UTEP, El Paso, TX 79968-0520 (915)747-6934 arrowood-at-utep.edu
} From a practical standpoint, wouldn't it be best to focus using the same conditions that are later used for taking the images, unless there are intensity or other issues that prevent that?
A few years back, when I was doing hi-res TEM, we would carefully set the conditions on the TV screen, then turn off the room fan, move away from the column, hold our breath and take a picture.
However, this may have been peculiar to hi-res materials work, where even tiny changes in focus can have dramatic effects.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Thursday, March 01, 2001 2:30 PM To: 'mlynn-at-miami.edu'; 'Microscopy-at-MSA.Microscopy.com'
In terms of contrast transfer theory (i.e. when linear imaging allows approximating the imaging process as convolution with a point spread function), the lens transfer functon doesn't depend on convergence. So for example you shouldn't effect the defocus of the image by slight changes of the illumination convergence.
However, the convergence imposes a damping on the total contrast transfer, which gets more severe when convergence is greater. For details on this 'spatial coherence envelope' see any text on HREM. Because of this, the image quality will improve if you record with a less condensed beam (better coherence).
With a field emission gun, you won't be able to image at crossover, because the source will almost certainly be too small - these guns give the most coherent illumination available.
For a thermionic gun, the limiting factor will be: if the convergence is too great at crossover, you won't see much because everything will be blurred out by the coherence limitation of the incident irradiation. If this is the case, you should either change condenser apertures to decrease convergence (then you can focus, and/or stigmate at crossover), or spread the beam to improve coherence.
As far as I can see, there is no reason why one should or should not adjust or even record images at crossover (except that if the filament is a bit undersaturated, you will get uneven illumination). Getting the best possible coherence is important, but it doesn't really matter how one achieves this optically. Decreasing intensity is therefore good, to the extent that sample drift doesn't become too much a limiting factor.
Overfocusing the illumination rather than underfocusing is better, I would say (based on experience). I'm not sure why - perhaps some of the energy spread gets filtered out more effectively - at any rate for the same size beam one has more intensity underfocused than overfocused, so something is being cut out by overfocusing.
Wharton
} -----Original Message----- } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } Sent: Thursday, March 01, 2001 2:30 PM } To: 'Microscopy-at-MSA.Microscopy.com' } Subject: RE: Focusing } } ------------------------------------------------------------------------ } 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 always "heard" to focus the image with the beam in an overfocus } condition } (past crossover) for improved beam coherence. I find it difficult to } focus at } crossover anyway. Also, I don't think you can accurately assess focus or } astigmatism with the beam at intensities suitable for most photography. } Depends on the magnification and a host of other things, of course, such } as } your exposure time. I don't think there's any justification to focus at } exactly the same intensity at which you will photograph. I too await } further } enlightenment from the list.... } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } [SMTP:TindallR-at-missouri.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 } } -----------------------------------------------------------------------. } } } } } } Hi all, } } } } I have one of those very basic "thought" questions on a procedure that } I've } } always taken for granted, but am now not so sure about. I had been } trained } } from Day One to always focus a TEM with the beam at crossover, then to } } spread the beam until the exposure intensity is reached. The idea seems } to } } be that small focusing errors at crossover will be corrected as the beam } is } } spread and "depth of focus" is increased somehow. } } } } The reason I'm asking is that in recent years, almost no one else I've } run } } across focuses in this way, preferring instead to focus with the beam } } already spread to the point at which the photograph will be taken (at } least } } as long as the image is bright enough to see for focusing). Out of } } curiosity, I ran tests using both methods and have noted no clear } } differences. } } } } Does anyone have any thoughts on this? Any ideas or theories about why } one } } method would be superior to the other? } } } } Sincerely, } } Curious in Columbia } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } }
*************************************************** PROGAM: FOCUS ON MICROSCOPY 2001 ***************************************************
Dear colleagues,
The program of the conference "Focus on Microscopy 2001", April 1 - 4, 2001, Amsterdam, has been finalised and can be found on our Web-site
www.focusonmicroscopy.org
This conference is the 14th in a successful series of interdisciplinary meetings on 3D acquisition and 3D image processing and forms an effective meeting point for both developers and users of 3D-microscopy. The more than 120 contributions this year range from new developments in Coherent Anti-Stokes Raman Microscopy (CARS) and multi-photon excitation to the application of such techniques in biology, medicine and material sciences. This year "3D Microscopy of living cells and tissues" will receive special attention, which reflects the growing number of researchers that use GFP-techniques and life-cell imaging for their experiments.
A substantial instrument exposition including the main manufacturers in the field will accompany the conference.
Please, visit our web site http://www.focusonmicroscopy.org for further details concerning the program and the conference.
Looking forward to seeing you in 1st of April in Amsterdam, on behalf of the program committee,
We are working with embbeded cells in LR White with and without osmium. (Polimerization was acomplished at 60ºC). Our purpose is to detect gangliosides (GD3). Do you have any suggestion?
I also was told early on in my career that one should focus at the magnification the picture was to be taken at, though I find it difficult to do this in practice because the illumination is often too low. I usually compromise and focus with as little illumination as I can get away with.
I wonder whether the reason for the change in focus is not due to changes in the illumination, but changes in the specimen position. At different beam densities the degree of specimen charging may vary, thereby changing the position of the specimen in the lens field. This might explain why some microscopes are less sensitive to the specimen current density than others. Certainly the amount of specimen charging seems to be affected by the objective aperture, for example.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
Peggy: I don't know if you received any responses off-line, and I hope that Ronald Austin was able to provide you with a protocol for his microwave procedure (actually, if you have one available Ronald, I'd like a copy, please). Your question is most appropriate because that is what I am (frantically) in the midst of doing. Altho' "frantic" applies only to the deadline.... Sans a microwave procedure, I am doing the old time-tested methods. The only thing I make sure of is the buffer used to transfer the tissue from formalin. I add 6% sucrose to the buffers and carry that through until I get into osmium. Loss of membranes occurs in the formalin, and if you add the 6% sucrose it seems to help retain membranes in the final EM sample.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On 26 Feb 2001 15:06:20 -0500 (EST), "HARGER-ALLEN.MARGARET_J-at-INDIANAPOLIS.VA.GOV"-at-sparc5.microscopy.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 | -----------------------------------------------------------------------. | | | Pathology lab. listers: are there any new/and/or superior techniques | for processing previously formalin fixed tissue for TEM? | Thanks, Peggy Harger-Allen | |TAB|email:harger-allen-at-indianapolis.va.gov |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
The pellets are from a hydrobiid snail found in highly mineralized warm desert springs and Posos in Mexico. I am interested primarily in the bacteria found on and in them but would also like to be able to inventory the entire contents of the pellets to help get a better picture of what part they play in their isolated ecosystem. I've done some light and SEM on samples but need to get inside to determine if the bacteria that break them down are carried internally or are picked up environmentally from the water. For sample preparation and sectioning I need to consider other contents like diatoms and stromatolite pieces and the high concentration of mineral solids in the samples. To start I plan to fix with glut/OsO4, dehydrate with acetone, infiltrate with propylene oxide and go into embed 812. After sections are imaged for non organic I planned on staining with lead citrate and comparing. Being new to TEM and not having a good idea of the precise osmolarity of the sample has left me puzzling over buffer selection (sodium cacodylate vs. phosphate), concentration, and times. We have an excellent EM technician here at NAU but it seems more my responsibility to make these determinations so any suggestions or references you could offer would be greatly appreciated. Thanks Pete Polsgrove Northern Arizona University pjp6-at-dana.nau.edu
we have just bought a Nikon CoolPix 990. It is an outstanding camera. One thing that we are disapointed it is a focusing problem when taking picture in Microscope. For any zoom setting and even in a manual mode, the micrograph borders always get out of focus, and in some situation with a Chromatic aberration. Does anyone have any idea how to eliminate this problem? The camera came with all necessary accessories for mounting in a microscope, e.g., c-mount, and MDC relay lens and remote cable.
Best regards,
Leonardo -- --- Leonardo Lagoeiro Departamento de Geologia Universidade Federal de Ouro Preto Ouro Preto, MG, 35400-000 Brazil E-mai: lagoeiro-at-degeo.ufop.br
The CoolPix 990 is indeed a nice camera. Unfortunately, it is not ideal for microscopy. It will work in this type of application but the results are mixed and the effort can be great.
If you zoom out to infinity, you should get best results. The key problem I have found is obtaining consistent exposure values. Preview with the puck looks good but the captured image is over exposed. Trying the best of five sometimes helps. But i have relegated the 990 to travel and snapshots. I don't think that it is ready from prime time microscopy. YMMV.
gary g.
http://photoweb.net
At 01:35 PM 3/2/01, you wrote:
} Dear All, } } we have just bought a Nikon CoolPix 990. It is an outstanding camera. One } thing that we are disapointed it is a focusing problem when taking picture } in Microscope. For any zoom setting and even in a manual mode, the } micrograph borders always get out of focus, and in some situation with a } Chromatic aberration. } Does anyone have any idea how to eliminate this problem? The camera came } with all necessary accessories for mounting in a microscope, e.g., } c-mount, and MDC relay lens and remote cable. } } } Best regards, } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br
I am trying to distinguish between neurons and glia in cortical tissue that has been embedded in epoxy resin (LX112/NMA/DDSA/DMP-30). Does anyone know how to do this on semithin sections? Toluidine-Blue staining imparts some subtle differences in the appearance of chromatin in the nuclei of neurons vs. glia, but i am looking for a staining procedure that provides a more obvious color difference (without going all the way to immunohistochemistry).
It is my understanding also that KMnO4 will react with NMA (even if i disolve the resin out??), so those procedures are out, unless I can find a substitute for KMnO4.
let me make two observations and see if you agree:
1) The Coolpix (and any other camera like it) is originally designed for snapshots. That means, that the emphasis is on "nice" pictures under normal (outside or flash) conditions. This does not necessary mean, that one can take good scientific images. This may be the reason that you had problems with exposure time. The camera may take a weighted measurement to calculate exposure time, for example only from the middle. If you take a snapshot outside, the most important feature is likely to be found somewhere around the center of the image. So it's important to get that part right. That might not apply to photography on a microscope.
2) The quality of the picture is affected by the weakest link in the chain. My suspicion is, that the lens on the camera is far inferior to anything you put on a microscope. For example, we spend a lot of time (and money) to develop and make our own lenses for our TEM cameras. I don't see the point of spending thousands of Dollars on good microscope lenses and then have all that negated by an inferior lens on the camera.
One word to Nikon: I am not trying to knock this camera, it is probably a very good camera (I don't know it myself). My remarks are general and apply to other camera manufacturers (Kodak, Canon, Fuji, etc.). This was just a thread which mentioned the Nikon camera. Plus, all these ramblings are my personal opinion.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Friday, March 02, 2001 10:20 PM To: Leonardo Lagoeiro Cc: MSA listserver
The CoolPix 990 is indeed a nice camera. Unfortunately, it is not ideal for microscopy. It will work in this type of application but the results are mixed and the effort can be great.
If you zoom out to infinity, you should get best results. The key problem I have found is obtaining consistent exposure values. Preview with the puck looks good but the captured image is over exposed. Trying the best of five sometimes helps. But i have relegated the 990 to travel and snapshots. I don't think that it is ready from prime time microscopy. YMMV.
gary g.
http://photoweb.net
At 01:35 PM 3/2/01, you wrote:
} Dear All, } } we have just bought a Nikon CoolPix 990. It is an outstanding camera. One } thing that we are disapointed it is a focusing problem when taking picture } in Microscope. For any zoom setting and even in a manual mode, the } micrograph borders always get out of focus, and in some situation with a } Chromatic aberration. } Does anyone have any idea how to eliminate this problem? The camera came } with all necessary accessories for mounting in a microscope, e.g., } c-mount, and MDC relay lens and remote cable. } } } Best regards, } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br
Does anyone have a GaAs cathodoluminescence attachment in a SEM? This means a photomultiplier optimized to 8500A, an S1 photomultiplier. I would like to look at light emitted from a GaAs diode under power. Or does anyone know of a lab that does have GaAs cathodoluminescence as a SEM attachment.
I would like to thank those colleagues who kindly provided their advice and shared their experience with us regarding the problems of vibratome brain sectioning that I posted on the list server a while ago.
We now use double-edge feather blades that we purchased from Ted Pella, the quality of sections are much improved, also sometimes it seems helpful at low amplitude rather than high one.
Xinran Liu, M.D., Ph.D.
Center for Basic Neuroscience UT Southwestern Medical Center 6000 Harry Hines Blvd., NA4. 214A Dallas, TX 75390-9111
Arron: Try using Araldrite 502 for your resin. It does not contain NMA. This should eliminate your concern with using KMnO4. The mixture I use is a 1-1 Araldrite 502 and DDSA, mix well then add your DMP-30 and mix again. Don't worry about the air bubbles.
Ron Austin (Research Associate) Dept of Pathology LSU Medical Ct. Shreveport, LA rla-at-mindspring.com
-----Original Message----- } From: "PostMaster {rla-at-mindspring.com} "-at-sparc5.microscopy.com [mailto:"PostMaster {rla-at-mindspring.com} "-at-sparc5.microscopy.com] Sent: Saturday, March 03, 2001 2:43 PM To: rla-at-mindspring.com
-----Original Message----- } From: Ronald Austin [mailto:rla-at-mindspring.com] Sent: Saturday, March 03, 2001 2:41 PM To: Microscopy Society of America
Email: pjp6-at-dana.nau.edu Name: Pete Polsgrove School: Northern Arizona University
Question: I'm looking for an protocol for imaging fecal pellets. The pellets are from a hydrobiid snail found in highly mineralized warm desert springs and Posos in Mexico. I am interested primarily in the bacteria found on and in them but would also like to be able to inventory the entire contents of the pellets to help get a better picture of what part they play in their isolated ecosystem. I've done some light and SEM on samples but need to get inside to determine if the bacteria that break them down are carried internally or are picked up environmentally from the water. For sample preparation and sectioning I need to consider other contents like diatoms and stromatolite pieces and the high concentration of mineral solids in the samples. To start I plan to fix with glut/OsO4, dehydrate with acetone, infiltrate with propylene oxide and go into embed 812. After sections are imaged for non organic I planned on staining with lead citrate and comparing. Being new to TEM and not having a good idea of the precise osmolarity of the sample has left me puzzling over buffer selection (sodium cacodylate vs. phosphate), concentration, and times. We have an excellent EM technician here at NAU but it seems more my responsibility to make these determinations so any suggestions or references you could offer would be greatly appreciated. Thanks Pete Polsgrove
1) The camera is of course intended for general purpose photography. Snapshots, indoors and outdoors. It has a miniature flash which is good for about ten feet distance. For microscopy, of course the flash is turned off.
2)There are some macro zoom situations where the camera does a nice job. But for compound transmitted and reflected work, I have not found it to be satisfactory. Optem has made a high quality interface for the CoolPix which interfaces to Zeiss, Olympus and many other 'scopes. I have a common optical tube with C-mount to CoolPix for Axioskop and Olympus. I cannot fault this adapter system at all. But as you point out, the native CoolPix optics are not intended for discriminatory microscopy work. At about $700 for the Optem adapter, the CoolPix package with remote release and AC adapter is not cheap.
I recently took 765 pix with the CoolPix on a trip to Australia. It did a super job. I used two 128MB AVL CF modules. Some images were in highest JPEG resolution and some were in XGA fine. All print on an Epson Stylus Photo 2000P with outstanding results. And, of course, they go to the Web without any problem.
The Pixera Penguin 600CL is quite a different camera. It is highly suitable for microscopy, but also will do excellent macro work with an inexpensive C-mount zoom lens. Optem also makes the adapter for this camera to the Zeiss and Olympus microscopes. Same high quality and fine results. I have done multiple slices and stitched them together for a final TIFF file size of 110MB.
The Pixera is non-interpolated 5.8M pixels using their Diractor prism device. At 2776x2074 pixels, it generates a 17.5MB TIFF file. In 16-bit mode, the size is twice that. As I mentioned in a prior post, their software supports multiple capture averaging and integration. It also does variable sized spot metering and average metering. The CL version has Peltier cooling and exhibits exceptionally low thermal noise. Four frame averaging reduces noise even further.
The Pixera is 10-bits per color while the Nikon MX1200 is 8-bits per color. The word on the street is that the next release of the MX1200 will be 10-bits per color. Whether this is worth anything to a particular user is their own decision.
Being a professional Nikon shooter for many years, I am no longer an intransigent fan. In fact, I have dumped nearly all of my Nikon SLR equipment in favor of Contax. This of course has nothing to do with microscopy. But the point is that in my extended experience, Nikon is struggling to develop new products, which by specifications, leapfrog their competition--while in practice, they do not.
Try before buy is a good operative in this respect. I really don't care who makes what, as long as it is good stuff. Sorting through all the marketing hype is a chore. Sometimes even working with the equipment is a chore. But the actual results of using products is much more telling than sales pitches or product brochures.
gary g.
http://photoweb.net
At 06:54 AM 3/3/01, you wrote:
} Gary, } } let me make two observations and see if you agree: } } 1) The Coolpix (and any other camera like it) is originally designed for } snapshots. That means, that the emphasis is on "nice" pictures under normal } (outside or flash) conditions. This does not necessary mean, that one can } take good scientific images. This may be the reason that you had problems } with exposure time. The camera may take a weighted measurement to calculate } exposure time, for example only from the middle. If you take a snapshot } outside, the most important feature is likely to be found somewhere around } the center of the image. So it's important to get that part right. That } might not apply to photography on a microscope. } } 2) The quality of the picture is affected by the weakest link in the chain. } My suspicion is, that the lens on the camera is far inferior to anything you } put on a microscope. For example, we spend a lot of time (and money) to } develop and make our own lenses for our TEM cameras. I don't see the point } of spending thousands of Dollars on good microscope lenses and then have all } that negated by an inferior lens on the camera. } } One word to Nikon: I am not trying to knock this camera, it is probably a } very good camera (I don't know it myself). My remarks are general and apply } to other camera manufacturers (Kodak, Canon, Fuji, etc.). This was just a } thread which mentioned the Nikon camera. Plus, all these ramblings are my } personal opinion. } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Friday, March 02, 2001 10:20 PM } To: Leonardo Lagoeiro } Cc: MSA listserver } Subject: Re: Nikon CoolPix 990 } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } } } Subject: } } } } SEM views of gold paint } } } } Content: } } } } Does anyone have any views of modern gold flake (real gold, not brass } } flake) used in making paintable substitutes for gold leaf? } } } } John Twilley } } Conservation Scientist }
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The candidate will be working in a diverse industrial R&D-environment with several links to university labs. He/she will be involved in a multidisciplinary team working on several nanostructured applications. Main focus of the work will be on microscopical and crystallographic (diffraction) techniques. In order to meet the technical requirements, the candidate should have a PhD in materials science, experience with TEM- and diffraction-techniques, and have a background in crystallography of inorganic materials. He/she should be a good teamplayer to function in a multi-disciplinary team, and be fluent in English and/or Dutch.
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For selection procedures, eligibility criteria and so on, please see homepage Marie Curie Industry Host Fellowships: http://www.cordis.lu/improving. Please note in particular that candidate fellows must be nationals of an EU Member or Associated State, or have resided in the EU for at least five years immdiately prior to their selection by Agfa.
I don't want to wake up a recent discussion (I was away in the last weeks), but I take advantage of this subject to askh a question about an other aspect of this subject. Sometimes you have so old material and maintenance services are laughing when they see what your are working with and/or on the other hand (with new materials also) it's faster and cheaper to do cleaning and (basic) maintenance yourself (and I like to do it myself !).
My question : What kind of grease/oil, where, on a LM ?
Manufacterer and maintenance services don't like to answer such a question. They say its complicate, there are much type of greases etc etc... So, does someone know something about that.
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
I was quite surprised to see the problems that are being discussed in relation to TEM focus, particularly as I was with Hitachi in the HU11 days, lets keep it simple and try to help?
When setting the image focus you should be aware of a number of problems that may complicate matters.
1. To try to focus at condenser cross over is absolutely the wrong thing to do! i) At crossover there is the poorest illumination coherence, this will lead to soft images, not out of focus but not optimised either. ii) At crossover there is a space charge effect (too many electrons in a small area and they repel each other) which may cause a false focus. (Just look at any image at crossover and then overfocus the condenser - you can see more detail!) iii) To focus at crossover and then change to a lower intensity is asking for trouble as the change in heating effect is likely to cause specimen drift and flexing which may change the contrast of material science specimens.
2. You MUST focus at the photographic magnification. Focus is the matching of the focal lengths of the objective and diffraction lenses. If you focus at a higher level and there is a diffraction lens change when dropping the magnification, you will find a matching objective correction will be required or your image will be out of focus.
3. What is best FOCUS, not always true focus? To a materials scientist it will almost certainly be true focus, no Fresnel fringes, as determined by the focus wobbler. They will set their intensity in advance as an intensity change may cause the material to flex and change in contrast. To the biological scientist it will be a degree of underfocus determined by the specimen's organelle density, its thickness, the kV, the objective aperture size and most important, the magnification; this is optimum under focus, the defocus where the (white) Fresnel fringes enhance the high contrast areas!
When taking a photograph the ideal conditions (as used in all the manufacturer's demo laboratories that I have worked with, Hitachi, JEOL and ISI) require the photograph to be taken at the same intensity level as when the focus is set. WHY? Well, once an intensity is set the operator may focus and during the focus procedure they should be concentrating sufficient to spot any image drift; it is most important to see the image under the conditions that you are going to photograph. So what will the excuses be?
1. "It is too dim to focus" - then i) increase the emission current, many people run at too low a current and make the task of optimising the instrument settings much more difficult ii) increase the kV which would increase the intensity too ii) re calibrate the photographic system to allow a focus intensity that is suitable for most operators
2. "The negatives are to dark" - i) the denser the negative the higher the contrast, so may be you could increase the kV, have a more stable specimen and an even brighter image?
3. Difficult one this - "the pathologists like a very bright image" - yes I know I guess it comes from looking down light microscopes all day? i) try 1 or 2 above - "yes but the pathologists like a high screen contrast" - yes I know but you should explain that no one publishes the screen, it it the photographic record that is the most important item(?) and we can get stacks of contrast from modern printing/publishing systems.
Regarding the reported overlap in currents between condenser and objective it is true, however the result is usually more of an image shift than a focal change in my observations, but yet another reason to focus and stigmate at the photographic intensity.
So to conclude I would
1. Always focus and stigmate at the photographic illumination level, if a biologist determine and plot out the optimum under focus for your material over your normal magnification range. 2. Always use the second condenser overfocus from crossover for high coherence and combine this with a 2 micron spot size for biology. 3. Set up the instrument's photographic procedure to suit as many operators as possible so that they too may focus at the photographic level. 4. Increase the emission current to give a higher brightness making point 2 far easier to use 5. Always use a higher kV (if 60, wow please go to 80, if 80 try 100) and work to obtain the best results on a photograph not trying to optimise for the screen. 6. To focus at low magnifications try removing the objective aperture and focus without a binocular, or a wobbler, looking for the very strong minimum contrast effect.
Well I hope this helps, its standard information on our courses, guess we do too much SEM in the States!
Regards
Steve Chapman Senior Consultant Protrain For consultancy and training by professionals World Wide Tel +44 1280 814774 Fax +44 1280 814007 www.emcourses.com
} } I have several questions for an AFM microscopist; } } 1. Can AFM really measure surface modulus?
Probably - but there are a number of factors which may have to be taken into account. These will include the nature of sample, any additional interactions between the sample and the scanning probe tip, the nature of the tip and SPM cantilever and so on.
} } 2. If yes, what is the mode of operation or measurement? } In terms of imaging modes (i.e. those where you are obtaining an image along with surface property information) there are two that may provide some surface modulus information.
i) Phase imaging (see for e.g.http://www.di.com/AppNotes/Phase/PhaseMain.html) is capable of providing a surface map based on the viscoelastic properties of the surface. Therefore the image will probably be a convolution between both the surface modulus and other surface properties (for example elasticity and whether the sample has any interactions with the tip). In this mode the probe is tapped into the surface and the lag between the voltage driving the tip oscillation and the actual tip oscillation provides this information. (It is particularly difficult, if not impossible to quantise the results obtained).
ii) Pulsed force should be able to deconvolute between sample stiffness and sample adhesion. http://www.thermomicro.com/tech/modes/pfm.htm should provide more information about this mode. However whether true values could be measured would again be a matter of some discussion.
Further mode of scanning probe microscopes are the spectroscopic ones in which the probe tip is move towards and away from the sample and the tip response measured. In AFM this mode is force-distance (see for e.g. http://www.thermomicro.com/tech/modes/fvsd.htm). This should again provide some information regarding the surface modulus. And with some tip calibration may provide something akin to a quantitative measurement.
With all of these mode the cantilever type will have an effect on the amount of information available. If the cantilever is less stiff than the surface of interest your measurement will generally be of the Young's modulus of the cantilever.
Nanoindenters may provide a more accurate measurement. http://freespace.virgin.net/micro.materials/ is an example. There are some companies that offer methods of converting commercial SPMs towards a nanoindenter.
} 3. Is it a quantitative measurement? Is it an absolute measurement } or a relative measurement?
My cynical opinion would be that most measurements available from a more qualitative than quantitative, though many would disagree. For example in this case, the area of tip-sample contact is unlikely to be able to be measured accurately but will have a major effect on your measurements, You may however be able to obtain some numbers that can be converted to an absolute measurement after careful consideration.
} } 4. Can the results be correlated to the (bulk) modulus measured by } rheometer?
This would, I presume, depend upon the material.
I hope this is of some use.
Giles
---------------------------------------------------------------------------- --------------------------- Dr. Giles Sanders Zeneca / SmithKline Beecham Centre for Analytical Sciences Chemistry Department Imperial College of Science, Technology and Medicine London SW7 2AY
I have had a request from a customer to allow remote access to our SEM in operation ( Hitachi S3500N ). The software they are proposing to use is "Timbuktoo" (?). They wish to set up three-way access across the internet along with a web-cam. I would appreciate advice from anyone who has tried this already. It would be useful to know what software has been used elsewhere and what problems have arisen. There may be better software available to carry out this function, but I have not approached this problem until now and am a bit in the dark. All advice, comments, etc. will be much appreciated.
Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
Dear Colleagues We have a CM200 (S)TEM equipped with EDAX IIs, 4pi digital imaging system and Gatan GIF. We are considering buying an integrated data acquision system for this microscope. ES Vision System from Emispec Systems, Inc. is under consideration . Comments from users of this ES Vision System would be highly appreciated. Of particular interest is operation of the Gatan Image Filter; would the autofilter functions still be available? Are there any other such systems out there? TIA Anita
To the person who is polymerizing the L.R. White at 60 degrees C:
Turn down the oven. If you want to do some sort of gold labeling on your sections try polymerizing the blocks at 45 degrees C for 2 to four days. Be sure to do this in some sort of BEEM capsule, as the stuff will not get hard if it is exposed to air. This type of plastic is not very stable in the electron beam, pick up your sections on carbon coated gold grids. Good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
First off, I suppose that Nestor or the folks at ORNL will have much more to say about this since they have essentially developed and refined telemicroscopy. However, I have some experience with Timbuktu that I will relate.
We have been using Tibuktu for a number of years since I first heard of it several years ago at an MSA workshop on telemicroscopy. It is designed for remote observation or control of computers in general and has been borrowed for the microscopy application. There are several other products also available for the same work. Norton has PC anywhere. ATT has developed VNC which has been mentioned here before and is free.
Timbuktu (and the others) simply requires a connection to the Internet. Whatever happens on the screen is potentially available to the world once a client logs in with a password. This can slow down the host computer as it also has to transmit a copy of the screen.
I don't know about PC Anywhere, but Timbuktu is much faster on the client side than is WinVNC, even on a LAN. There can be a lot of data to transmit and it flat out takes a while to do so. Well-written code can do a lot to speed that up. You should also consider the connection between your PC and your client. If they have only a modem connection, refresh rates will be painfully slow.
I couldn't guarantee how well Timbuktu (or any other program) would work on your scope. I seem to recall that there were some windows whose contents were not available for Timbuktu to share. It may be that the SEM control programs were doing non-standard operations that circumvented the normal Windows system. Presently, I am able to view video windows whether they be from Oxford's AutoBeam, Quartz's PCI or Dazzle's video preview. But don't expect refresh rates measured in frames per second, at least not with a remote control/viewer program. Still, it could be a good alternative compared to more expensive solutions or to traveling to the site.
BTW, we also picked Timbuktu since it worked with either Macs or Windows machines.
I cannot tell you much about web cams. I know some systems are available that are stand-alone and practically plug and play. I don't know that I would want to burden my microscopy or EDS computer with the extra task. Others may have more to say.
There, that ought to be worth a couple of cents.
Warren
At 03:57 PM 3/5/2001 +0000, you wrote: } Hi, } } I have had a request from a customer to allow remote access to our SEM in } operation ( Hitachi S3500N ). The software they are proposing to use is } "Timbuktoo" (?). They wish to set up three-way access across the internet } along with a web-cam. } I would appreciate advice from anyone who has tried this already. It would } be useful to know what software has been used elsewhere and what problems } have arisen. There may be better software available to carry out this } function, but I have not approached this problem until now and am a bit in } the dark. } All advice, comments, etc. will be much appreciated. } } Thanks. } } Best wishes, } } Colin
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Timbuktu is a remote control software package made by Netopia.
http://www.netopia.com/software/
There is a free trial version which can be downloaded.
Whether it will work for you, I don't know. One of the key issues is real-time image transfer for mag and focusing. The interconnect between the SEM and the remote user is a major factor in response time. Slow analog modems are not going to be all that responsive. If running on a LAN or intranet, that would make a big difference.
A similar product is PC-Anywhere.
gary g.
At 07:57 AM 3/5/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You can do it very cheaply (i.e. free if you have Windows machines) using NetMeeting. Our LEO 1530 was easily set up to do this. You can do "over the shoulder" microscopy where the remote location observes. You need fast bandwidth for control and the instrument must be fully digitally controlled. We have tried it out with remote locations within our company and it works, but do not use it very much (i.e. never).
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center 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)
-----Original Message----- From: Colin Reid [mailto:creid-at-truxa1.tcd.ie] Sent: Monday, March 05, 2001 10:57 AM To: MSA.Microscopy.Com (E-mail) Subject: Remote Access to SEM
--------------------------------------------------------------- --------- 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,
I have had a request from a customer to allow remote access to our SEM in operation ( Hitachi S3500N ). The software they are proposing to use is "Timbuktoo" (?). They wish to set up three-way access across the internet along with a web-cam. I would appreciate advice from anyone who has tried this already. It would be useful to know what software has been used elsewhere and what problems have arisen. There may be better software available to carry out this function, but I have not approached this problem until now and am a bit in the dark. All advice, comments, etc. will be much appreciated.
Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
to make a guess, you should can give some more information. What impurity elements did you find/expect ? Can you make any suggestion about the size of any kind of inhomogeneity - conzentration, precipitation or what ever ?
S.Guder
} We're analysing Cu wire here for purposes better known to ourselves. We } are noticing, but not able to image interesting variations in the } concentration of impurities, which suggests to us, either that the } original material was inhomogeneous, or that drawing the wire has caused } some kind of exsolution process. Have any of you encountered smilar } things in metals? Refs? } Malc. } } -- } Dr MP Roberts Phone: [+27](0)46 603 8313 } Dept of Geology Fax: [+27](0)46 622 9715 } Rhodes University Cell: 083 4060 262 (usually off) } 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za } SOUTH AFRICA
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr.-Ing. Susanne Guder Technical University Munich Department of Materials in Mechanical Engineering D - 85747 Garching Phone: + 49-(0)89-289-15308/15338 Fax: + 49-(0)89-289-15301 Email: guder-at-wm.mw.tum.de ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I own a zeiss 63X water immersion lens. If the coverslip thickness is 0, does that mean that this is meant for direct viewing of specimens in water? I purchased it with that in mind (I work in marine environments, felt it would be convenient to observe microorganisms and microanimals directly). Using a coverslip doesn't seem to work well, if at all. For some specimens it is nice, but any focusing movement will push specimens out of the way. Nice for zooxanthellae in a small sea anemone, for example, and was pretty good for a thick broth of unicellular green algae.
I am trying to get a feel for this objective. WOuld appreciate any information possible.
Alan Davis adavis-at-saipan.com
On Fri, 13 Oct 2000 08:51:07 -0400 Gary Radice {gradice-at-richmond.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've had some experience with water immersion lenses. Their main } advantage is higher numerical aperture than dry lenses at } equivalent } magnification, so better theoretical resolution. They don't offer } quite as good resolution as oil immersion lenses, but they tend to } have longer working distances, I believe, and don't have the } messy } clean-up of oil immersion lenses. So, yes, they do have some } distinct } advantages. } } However, as others have pointed out, you may already be able to } see } everything you need to see with your current lenses. And nothing } beats a test-drive. } -- } Gary P. Radice gradice-at-richmond.edu } Associate Professor of Biology 804 289 8107 (voice) } University of Richmond 804 289 8233 (FAX) } Richmond VA 23173 http://www.science.richmond.edu/~radice
-- adavis-at-saipan.com 1-670-235-6580 Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, NMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
Has anyone priced the XL30 high vac FESEM and the EFESEM or variations which use LaB6? What is the general price range for these, and which Hitachi models would closely compete?
A normal (upright) epifluorescence microscope excites fluorescence in a specimen by passing the excitation light to the top of the specimen via the objective lens, which is used to focus the light into an intense spot exactly at the point on the specimen which is to be viewed. Fluoresced light from the region of interest is collected by the objective and is viewed or photographed normally after filtration to remove any excitation light and unwanted fluorescence wavelengths. The excitation illumination is commonly provided by a high-pressure mercury vapour lamp, which conveniently provides very high intensity illumination in UV, blue and green wavelengths. Other high-intensity sources are now also used, including xenon lamps, lasers (e.g. in confocal microscopes, which are a highly-derived type of epifluorescence microscope) and even tungsten-halogen lamps. The separation of excitation and fluorescence wavelengths is usually accomplished using a dichroic beam splitter which reflects green, for example, and transmits orange/red. Further fine-tuning of the excitation wavelengths may be done using dichroic filters. Many microscope manufacturers provide filter systems as beam-splitter cube assemblies with all the filters and dichroic reflectors required for a particular fluorochrome installed in one pre-aligned package for easy exchange. The fluoresced light may be further filtered for viewing or photography by "barrier" filters either of dichroic type, or of coloured glass or sometimes gelatin (e.g. Kodak Wratten).
Other than that, an epifluorescence microscope can be a normal compound microscope, but because efficiency of illumination and collection of light is important, and is limited by the objective lens, fluorescence microscopes usually employ high quality objectives with very high numerical aperture. If UV is used, it may be necessary to select ojectives with very low UV absorption.
Hope this helps Chris Date sent: Mon, 5 Mar 2001 17:25:50 -0600 To: Microscopy-at-sparc5.microscopy.com } From: ileyozerlat-at-yahoo.com ()
The FEI XL30SFEG costs in the region of £250k in UK The closest Hitachi competitor is the S4700. The FEI is thermal field emission, the Hitachi is Cold cathode field emission. There is no Hitachi ESEM, but they do a variable pressure tungsten filament SEM.
Philips / FEI service in UK is excellent. I have no first hand knowledge of the Hitachi service operation, but believe it is also first rate. Hitachi quote much lower prices for their service contracts than FEI. They tell me that their engineers generally have little to do because the instruments are so reliable! I don't know whether this is a true reflection of the cost of ownership, however.
Date sent: Tue, 06 Mar 2001 02:03:30 -0800 To: MSA listserver {Microscopy-at-sparc5.microscopy.com} } From: Gary Gaugler {gary-at-gaugler.com}
Hello,
Our Siemens Elmisjop 102 is presently out of order. We are looking for several parts of a Siemens Elmiskop 102 which must be replaced.
Parts List C73000 - E3174 - C1 - 1
Part N°335 - Objective current control fine / superfine - cat n° C72315 - A29 - A4
Part N°333 - Objective current control coarse I level - cat n° C72315 - A31 - B1
Part N°334 - Objective current control medium II level - cat n° C72315 - A31 - B2
Part N°455 - P.C. board V9 "Objective controller" LR - cat n° C72302 - A41 - A2
Many thanks in advance
Best regards
Philippe Drouillon Solvay Research and Technology Electron Microscopy and Image Analysis - Coordinateur Informatique de Division Rue de Ransbeek, 310 B-1120 Brussels (Belgium) phone : (00 32) 2 264 24 47 mailto:philippe.drouillon-at-solvay.com
We have an Emispec Vision system on our CM200FEG TEM-STEM to acquire spectrum lines and spectrum images from our Oxford EDS and Gatan GIF. We tune the GIF using the autofilter functions, and setup the GIF with ImageFilterControl, but let ESVision read out the GIF multiscan camera during acquisition. All in all, we are quite pleased with the system. If you would like to discuss this offline, please contact me directly.
} Hi, } } I am trying to distinguish between neurons and glia in } cortical tissue that has been embedded in epoxy resin } (LX112/NMA/DDSA/DMP-30). Does anyone know how to do } this on semithin sections? Toluidine-Blue staining } imparts some subtle differences in the appearance of } chromatin in the nuclei of neurons vs. glia, but i am } looking for a staining procedure that provides a more } obvious color difference (without going all the way to } immunohistochemistry).
I think you are going to have to rely on morphology alone. Get a copy of Peters, Palay and Webster, "Fine Structure of the Nervous System". You should be able to extrapolate from the low mag EMs to LM. You could probably do an immuno for GFAP (most but not all astrocytes) but that still leaves oligos and microglia. I don't know of a good immuno for these cells on plastic sections. Plus, there will be smaller neurons to contend with. Sorry!
} It is my understanding also that KMnO4 will react with } NMA (even if i disolve the resin out??), so those } procedures are out, unless I can find a substitute for } KMnO4.
And the KMnO4 is for ??
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 mcauliff-at-umdnj.edu **********************************************
There is an official open position now at Genentech (the one I alluded to in an earlier post, but HR is not too speedy over there) . The posting is below and can also be found on their website: http:// www.genentech.com {http://www.genentech.com/} .
For immediate consideration please send a c.v. to Peter Schow at pschow-at-gene.com {mailto:pschow-at-gene.com} . He is not on the list, so please email him directly. I have already forwarded previous resumes I received to him, so no need to resend those.
Position: Research Associate- Cytometry Lab Requisition #: 01-0003250
Description Research Associate position available in the Research Cytometry Lab. The lab support core facility, collaborative and basic research functions. Primary responsibilities include operation and maintenance of flow cytometers and/or imaging instrumentation and may cover all aspects of data acquisition, analysis, interpretation and presentation in the contexts of basic and applied research. Lab supports Coulter and BDIS flow cytometers, Leica (confocal) and Nikon (digital fluorescence) microscopes and both Mac and PC computers. The person will work in a dynamic and interactive environment requiring a strong teamwork orientation, good communication skills, flexibility and the ability to interact with a diversity of research personnel.
Requirements Position requires a BS/MS and extensive experience with flow and/or image cytometry. Good working knowledge of cell and molecular biology necessary. Industrial experience is desirable.
Holly Aaron Head, Berkeley Imaging Center University of California, Berkeley Dept. of Molecular and Cell Biology 447 LSA Berkeley, CA 94720-2751 hollya-at-socrates.berkeley.edu
Northwestern University is drafting a complaint against a vendor of microscopy products, AMT. Has anyone else had experience with lawsuits against vendors?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
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Nestor Your Friendly Neighborhood SysOp
=========================================== Dr. Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
I am considering this, but have read about issues of contrast perception and/or low light vision. It occurred to me that microscopists, particularly EM types, depend on their eyes in low-light situations more than any other profession I can think of, with the possible exception of cat burglars.
Has anyone out there had LASIK or similar procedure, and has it been positive or negative for your work? Did you notice reduced contrast sensitivity?
Thanks for any personal experiences; since it's not a direct microscopy query, please send replies to me and (if there's interest) I'll summarize for the list.
Rick Mott (myopic, -7 or so, with some astigmatism)
We have a ten year old cryo-stage and transfer device for sale.
It is a BioRad E7400 comprising: Pump valve controller Sputtering module Evaporation module Edwards E2M5 rotary pump x2 Valve block Transfer/coating unit with dewar Stage cooling unit with dewar Specimen insertion rod ie all you need to go cryo-SEMing.
The rod and flanges suit a JEOL 6400 SEM. The unit was functioning and in good condition prior to replacement. Any offers?
Eric Hines Microscopy Centre CSIRO Entomology Canberra.
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
It should be possible to do this much faster and without any sample preparation with X ray reflectometry.
See web site :
http://www-cxro.lbl.gov/optical_constants/
The is there the possibility to simulate some mesures.
Optical methodes could also be able to do this, by interferences in the TiO2 film.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
we are interested in purchasing a good software for desktop- analysis (on a PC) of X-ray EDS spectra, previously acquired on an analytical TEM and transferred to other computer.
In the book of Williams and Carter is mentioned only one such program, i.e. the DTSA, from NIST. Does anyone of you know a web address where I could get more informations about this software ? I would be delighted if there will be any possibility to check it before ordering, but where or how? Besides, does anyone know about a better program of this kind ? We would much better appreciate a free one, but we are ready to purchase even a commercial one.
Thank you for your attention.
Corneliu Sarbu, PhD Metallyrgy and Applied Materials Science Dept. Catholic University of Leuven Belgium
---------- } From: "CAROL.A.WALDER" {eencaw-at-elec-eng.leeds.ac.uk} To: mtlrmdb-at-leeds.ac.uk
Hi Listers,
I have a LKB Bromma 11800 Pyramitome whose belt has come loos and it needs to be replaced. I can't find the instruction manual or the slip of paper that my predecessor wrote the service person's name on.
Does anybody out there have a source for the instruction manual? I called Leica but they told me this machine was absolete when they acquired the LKB line.
I'll fix this thing myself if anyone can send me a copy of the manual (I'll glaly pay for the cost of copying and shipping).
If anyone knows of a repair person who still works on these ancient beasties, please let me know. It would be good to have someone who could give them tune-ups every now & then.
My screws are loose and I've got the hammer, blow torch sosme bubble gum and bailing wire....now all I need is the manual.
Pretty please??
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
An excellent write-up about the process is located at http://www.manufacturing.net/magazine/dn/archives/current/feature1.html
Cheers Peter Tarquinio peter-at-evex.com
Evex Analytical Microanalysis and Digital Imaging 857 State Road Princeton, NJ 08540 609-252-9192 T 609-252-9091 F www.evex.com info-at-evex.com ----- Original Message ----- } From: "Rick Mott" {rickmott-at-alumni.princeton.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, March 06, 2001 11:20 PM
} In the book of Williams and Carter is mentioned only one such } program, i.e. the DTSA, from NIST. Does anyone of you know } a web address where I could get more informations about this } software ? I would be delighted if there will be any possibility to } check it before ordering, but where or how? } Besides, does anyone know about a better program of this kind ? } We would much better appreciate a free one, but we are ready to } purchase even a commercial one. } } Thank you for your attention. } } Corneliu Sarbu, PhD } Metallyrgy and Applied Materials Science Dept. } Catholic University of Leuven } Belgium
Corneliu,
DTSA is no longer sold as a Standard Reference Data product. NIST has abrogated its patent on DTSA so both the executable and source code are available from the following URL free of charge:
If you find it does not meet your needs, please drop me an email. Our group welcomes suggestions for new features or improvements.
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
You can find information about DTSA at the NIST web site
http://www.cstl.nist.gov/div837/837.02/dtsa.html
There is only a Macintosh version of this software, so it may not be of use for you if you are looking for software for a PC. NIST has officially withdrawn DTSA as a Standard Reference Data product and has abrogated its patent. NIST is now releasing the software free from charge, however no longer officially supports the software except for internal development as an internal research tool for NIST. The NIST web site I directed you to has links to download the software application and the Pascal source code.
Tyrone Daulton
} Dear fellows microscopists,
} ... } we are interested in purchasing a good software for desktop- } analysis (on a PC) of X-ray EDS spectra, previously acquired } on an analytical TEM and transferred to other computer.
} In the book of Williams and Carter is mentioned only one such } program, i.e. the DTSA, from NIST. Does anyone of you know } a web address where I could get more informations about this } software ? ...
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Experienced Histotechnologist to work in Neuropathology Research Laboratory of Department of Pathology
The person we seek will be responsible for maintaining a neuropathology research laboratory in the Department of Pathology at SUNY Downstate Medical Center. Knowledge of immunohistochemistry, special stains, and histopathology is required. Previous experience in handling central nervous system tissue preferred. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in a laboratory with hands-on experience in tissue processing, histochemistry, immunohistochemistry, and operation of a light microscope. Experience with technical aspects of neuropathology and performing special stains including silver (Bielschowsky) and myelin stains.
Two years experience working on immunohistochemistry of CNS tissue specimens with knowledge of immunoreagents and experimental protocols.
BachelorUs degree in science desirable.
Laboratory skills including communications, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, operation of computers and office equipment.
Advanced computer skills (word processing and database management) essential.
Desirable Experience
Confocal microscopy desirable.
Salary commensurate with experience
Responsibilities
Purchasing supplies and equipment, budget reports, laboratory maintenance and brain banking.
Will operate all microscopic, photographic and computer equipment, and keep accurate records of all laboratory experiments and procedures. Light and fluorescent microscopy; tissue processing for paraffin embedding, sectioning and slide stainer for immunohistochemical procedures; computer imaging (PhotoShop); general photography; and library and web searches
Familiarity with computer software for keeping records, report preparation and table/figure construction using Microsoft Office software (Word, Excel, PowerPoint) as well as Endnote and Adobe PhotoShop.
Send resume to:
Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
------------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu ------------------------------------------------------------------------
Experienced Confocal Microscopy/Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for organization of a new facility that includes a confocal microscope and an electron microscope. The position includes overall management of the microscopy facility, user training, and user supervision. Requirements for the position include experience with light and transmission electron microscopy. This individual will oversee all aspects of specimen accession and processing, operation of the microscopes, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Experience with confocal and digital imaging techniques, microinjection, visualization of living cells containing fluorescent probes, photobleaching, and fluorescence in situ hybridization.
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
Excellent interpersonal and organizational skills are essential.
BachelorUs degree in science desirable.
Desirable Experience
Expertise in training in the operation of confocal microscope systems is a distinct advantage.
Familiarity with light microscopy methods, immunofluorescent staining, use of fluorescent probes for physiologic measurements and the general principles of cell biological research are critical. Significant facility with computers is desired.
Responsibilities
Serve as the technical manager of the facility and be responsible for the operation and maintenance of the confocal and EM microscope facility.
Perform routine transmission EM, including tissue processing, ultramicrotomy, and examination; do preventative maintenance on the equipment; maintain the lab, order supplies, schedule instruments, and oversee billing.
Image analysis at the light, confocal and electron microscopic levels and preparation of micrographs for publication.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu
----------------------------------------------------------------------- Experienced Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for the overall operation of the EM laboratory in the Department of Pathology at SUNY Downstate Medical Center. This individual will oversee all aspects of specimen accession and processing, operation of the microscope, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
BachelorUs degree in science desirable.
Laboratory management skills including effective written/verbal communication skills to interact with a diverse group, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, and operation of computers and office equipment.
Desirable Experience
Previous experience in confocal microscopy highly desirable.
Previous experience in performing immunocytochemical staining and advanced computer skills usage (e.g. image analysis) is also desirable.
Salary commensurate with experience
Responsibilities
Maintain electron microscope in operating condition. Process clinical and research tissues for "thick" and "thin" sectioning. Darkroom management of photographic printing.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by: Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
{fontfamily} {param} Times {/param} {bigger} SCANNING ELECTRON MICROSCOPY (SEM) AND ENERGY DISPERSIVE
X-RAY SPECTROSCOPY (EDS)
WITH APPLICATIONS AT VARIOUS PRESSURES AND ATMOSPHERES
AN EDUCATIONAL SEMINAR-FRIDAY, APRIL 27, 2001
MOTOROLA GALVIN CENTER-SCHAUMBERG, IL
8:30 AM - 5:00 PM
{underline} Instructors {/underline} : Vern Robertson, JEOL-USA, and Nestor Zaluzec, Argonne National Laboratory.
{underline} Time {/underline} : Check-in at Galvin Center 8:30-9:00 am. Seminar 9:00-12:30 and 1:30-5:00.
{underline} Preregistration Required {/underline} : Deadline April 20, 2001; no walk-ins.
{underline} Cost {/underline} : Members of ASM-International and of the Midwest Microscopy and Microanalysis Society-$30; Non-members-$50; Students: $10. Registration includes lunch and refreshments at breaks. Check or money order to accompany preregistration form (below).
{underline} Exhibits {/underline} : Representatives from the following industrial co-sponsors will be present to discuss your applications and their products: EDAX, FEI, Hitachi, JEOL-USA, LEO, Noran, Oxford, and PGT.
{underline} Directions to Seminar Site {/underline} : Please see the other side.
{underline} Preregistration {/underline} : Please complete the following form. Mail it with your check or money order made out to "Chicago Regional Chapter of ASM-I" to Ms. Sheila Jungman, MSD 212, Argonne National Laboratory, Argonne, IL 60439 {bold} . {/bold} Deadline for receipt of the form is April 20, 2001. If you require a receipt for the preregistration cost, please so indicate below for pick-up at the Seminar Check-In desk. Questions? Phone 630-252-4157 or e-mail allen-at-aaem.amc.anl.gov.
{bold} SEM/EDS Educational Seminar-Friday, April 27, 2001
{/bold} Name (please print clearly)________________________________________________________
We are having problems with preparing brittle TEM sample of (single crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any suggestions beyond a kid-gloves approach?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Mary-Jacque, Several years ago at a used book library sale, I found a copy of Forensic Geology-Earth Sciences and Criminal Investigation by Raymond C. Murray and John C.F. Tedrow. It dates from 1975 and looks interesting. I hope to read more of it. Another title which came from a special session at the MAS-MSA meeting in 1985 is titled Electron Microscopy in Forensic, occupational and environmental health sciences by Samarendra Basu and James R. Millette. I still lament that the Microbeam Analysis Society dropped Sherlock Holmes' hat and pipe from their logo.
Lawry, I don't know about magnesium orthovanadate, but sapphire will work using the small angle cleavage technique. I have done cross sections of GaN on sapphire when I taught some students at Univ. of Illinois how to do it. John McCaffrey has done YBCO on cerium oxide on sapphire. If you are only interested in the bulk, that is even easier. If the magnesium orthovanadate is as brittle as you say, then SACT will probably work on that also. If you want, I can send you an image of the GaN/sapphire.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center 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)
} -----Original Message----- } From: L. D. Marks [mailto:ldm-at-risc4.numis.nwu.edu] } Sent: Wednesday, March 07, 2001 1:22 PM } To: Microscopy List } Subject: Brittle samples } } } -------------------------------------------------------------- } ---------- } 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/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } We are having problems with preparing brittle TEM sample of (single } crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any } suggestions beyond a kid-gloves approach? } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html } }
A quick clarification before I get deluged with responses about how to do cross-sections; we are not trying to do this. All (?) we want is a "conventional" 3mm disc sample of sapphire or magnesium orthovanadate. No glue, no mounting on a Cu ring, no nothing.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
I think a good solution may be Tripod Polishing. We have an application note on preparing samples of sapphire with a GaN film which may be useful. Let me know if you would like a copy of the note and I can email it to you.
Best regards-
David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
} RE: Brittle samples } ------------------------------------------------------------------------ } 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 having problems with preparing brittle TEM sample of (single } crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any } suggestions beyond a kid-gloves approach? } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } -------------------------------------------------------
Has anybody out there had any experiences with making your own transmitted electron detector for SEM? I'm interested in sharing ideas, references, etc. I've made one for myself and am fairly happy with its performance, but am wondering if there is more that I can do.
Thanks in advance,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I have two questions for you immunolabeling experts.
First, a client has some precious and irreplaceable histological sections of human hippocampus that have been labeled with ABC - DAB. Will it be possible for me to de-paraffinize the sections, expose them to osmium vapor and re-embed them in resin on the slide and pop them off and resection them and see the DAB precipitate? Anyone have a protocol that has worked?
Second, these researchers plan to use Vibratome sections of mouse brain and immunolabel them for light microscopy using an ABC kit. I have done on-ultrathin-section colloidal gold immunolabeling for TEM. However, for this new project we would like to try pre-embedding labeling of 50-60 micrometer Vibratome sections, then embed and resection them for TEM. Knowing what works for light microscopy, we'd like to use streptavidin/colloidal gold or whatever for TEM visualization. My question is, of course, does anyone have a favorite protocol they would be willing to share? How do I keep the sections from curling? If we stick them on a glass slide to embed in resin and (hopefully) pop off later for resectioning, when and how do I get the to adhere?
Mahalo!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
The URL I gave this morning for DTSA was correct, but not memorable. A few hours ago an alias was created to make it easier to navigate to the NIST Microanalysis Software web page. DTSA can now be accessed from:
http://www.nist.gov/dtsa
This page also contains links to Lispix, Dave Bright's excellent image processing application (now available for Win9x/WinNT/Win2k) and the NIST Monte Carlo codes. Lispix can be accessed directly at
http://www.nist.gov/lispix
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
Try epoxying a thin metal washer on one side of the sample, and dimple from the washer side.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
---------- From: L. D. Marks Sent: Wednesday, March 7, 2001 10:21 AM To: Microscopy List Subject: Brittle samples
------------------------------------------------------------------------ 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 having problems with preparing brittle TEM sample of (single crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any suggestions beyond a kid-gloves approach?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
The purpose of gluing a metal ring to brittle samples before dimpling is to support the sample. The sample is then waxed to the dimple grinding support stub and dimpled through the washer. We use this method routinely with brittle samples and not just cross sections. It's a good idea to carefullly clean the sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot washers usually work well for us, but we sometimes use other washer materials to avoid confusing Mo in the sample with Mo deposition artifacts.
Larry
---------- From: L. D. Marks Sent: Wednesday, March 7, 2001 11:38 AM To: Microscopy List Subject: Brittle samples - NOT CROSS-SECTION
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A quick clarification before I get deluged with responses about how to do cross-sections; we are not trying to do this. All (?) we want is a "conventional" 3mm disc sample of sapphire or magnesium orthovanadate. No glue, no mounting on a Cu ring, no nothing.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
the metallurgy or materials science group in your university should be able to tell you all about grain size measurements.
Best regards
Sam Purdy Technical Center National Steel Corp. Trenton Mi USA spurdy-at-nationalsteel.com
} ---------- } From: James Clarke } Sent: Wednesday, March 7, 2001 11:45 AM } To: NIH-IMAGE-at-LIST.NIH.GOV } Subject: countig grain size in archaeological iron samples } } IMAGERS! } Hope some one can point me in the right direction. I need to be } able to measure the grain size of metal samples which are of a } heterogenous nature. I am using a PC and Scion image seems to } have some of the things I need but some advice would be nice. } } --------------------------------- } James Clarke } J.Clarke1-at-bradford.ac.uk }
Fellow Microtomists: I am seeking information regarding 2 Sorvall MT2B ultramicrotomes that still are in use and in good working condition for both semi-thin sections and ultrathin sections. However, the faster speed of the return stroke of the cutting cycle no longer engages, even though the duration and position control belts are in tact for the slow-speed (cutting phase) of the cycle. Is this a relatively simple repair, and is there a company in the Cincinnati, Ohio, area that provides such repair service? [The serial numbers of these microtomes are 7701673 and 7800584. Do the first 2 digits reflect the year of manufacture (1977 and 1978 respectively)?] What would be a fair price for microtomes of this ventage? Thanks in advance for your assistance. EL Cardell
} } } Email: de-at-payload.com } Name: De McKeown } } Organization: Payload Systems } } Education: Graduate College } } Location: Cambridge, MA, USA } } Question: I need to determine the number of TV lines my microscope is } giving me. I have a test target but it only goes to 266 l/mm and I have } better resolution than that. I can't find a 'better' target and am stuck } trying to get an answer. How can I do this or where can I go for a new } target? } } I also have to find a contrast, but haven't got enough information yet to } ask a good question. } } Thank you!! }
There is a rash of virus mail out on the net again, a number of which are using "suggestive names" in the subject line. As a precaution some people trash messages which appear out of context.
While Humorous Titles are good for the soul (especially mine) and bring a smile especially when one gets the implied joke, let me remind you to at least add in the Subject Line with some standard notation so that individuals can more easily judge source of the mail. After all, we know that my Email filtering system is not perfect and suspect mail leaks through every so often.
Some Keywords include: TEM, SEM, EDS, .Confocal, LM, SamPrep, etc... check the FAQ site if you need idea's, but you can easily see what I mean.
For example: the posting on "I've got the screws loose" could have be preceeded or prepended by an appropriate Keyword like Microtome, even if the word Microtome was in () or abbreviated as uTome.
Unfortunately getting Mo (or anything else) on the sample destroys it as far as I am concerned. So, we have to rule out anything like this unless we can safely remove the metal with an acid (and not dissolve the oxide in the process).
On Wed, 7 Mar 2001, Thomas, Larry (PNNL) wrote:
} Laurie, } } The purpose of gluing a metal ring to brittle samples before dimpling is } to support the sample. The sample is then waxed to the dimple grinding support } stub and dimpled through the washer. We use this method routinely with brittle } samples and not just cross sections. It's a good idea to carefullly clean the } sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot } washers usually work well for us, but we sometimes use other washer materials to } avoid confusing Mo in the sample with Mo deposition artifacts. } } Larry } } ---------- } From: L. D. Marks } Sent: Wednesday, March 7, 2001 11:38 AM } To: Microscopy List } Subject: Brittle samples - NOT CROSS-SECTION } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } A quick clarification before I get deluged with responses about } how to do cross-sections; we are not trying to do this. All (?) } we want is a "conventional" 3mm disc sample of sapphire or } magnesium orthovanadate. No glue, no mounting on a Cu ring, } no nothing. } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html } } } }
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
A researcher has asked me to localize gluconase (a protein) in plant roots. They provided me with antibody (sera) and preimmune, which they have used in westerns. The problem is the preimmune is labelling the cell wall; the labeling is quite impressive and very specific. Now they tell me (alas I didn't ask before) that yes they have background on the westerns, but the preimmune never labels the 32 Kd protein they are interested in. What is the best approach to localize this 32 Kd protein? Should we cross absorb everything else out, or would it be better to affinity purify for the desired antibody. And why does this rabbit have such good antibodies to cell wall?
Thank you, Michelle
Michelle Peiffer ************************************************************* Electron Microscope Facility for the Life Sciences Penn State University Biotechnology Institute 001 South Frear Lab University Park PA 16802
The key to thinning brittle hard samples is to start with a doubly polished thin section 30 micrometers thick. Cut out 3 mm diameter discs with an ultrasonic probe. Glue these to a glass slide with crystalbond, a thermal plastic cement that dissolves in acetone. Mechanically thin and polish to less than 10 micrometers or thinner. Ion thin until satisfied.
Start thinning a thin sample. Don't break it. One good sample is all you need.
I have made many ion thinned samples of brittle materials, mainly shocked minerals, for over thirty years. Also this method is not for those in a hurry.
Gordon Nord USGS Emeritus Scientist
"L. D. Marks" 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 } -----------------------------------------------------------------------. } } Unfortunately getting Mo (or anything else) on the sample destroys it } as far as I am concerned. So, we have to rule out anything like this } unless we can safely remove the metal with an acid (and not dissolve } the oxide in the process). } } On Wed, 7 Mar 2001, Thomas, Larry (PNNL) wrote: } } } Laurie, } } } } The purpose of gluing a metal ring to brittle samples before dimpling is } } to support the sample. The sample is then waxed to the dimple grinding support } } stub and dimpled through the washer. We use this method routinely with brittle } } samples and not just cross sections. It's a good idea to carefullly clean the } } sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot } } washers usually work well for us, but we sometimes use other washer materials to } } avoid confusing Mo in the sample with Mo deposition artifacts. } } } } Larry } } } } ---------- } } From: L. D. Marks } } Sent: Wednesday, March 7, 2001 11:38 AM } } To: Microscopy List } } Subject: Brittle samples - NOT CROSS-SECTION } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } A quick clarification before I get deluged with responses about } } how to do cross-sections; we are not trying to do this. All (?) } } we want is a "conventional" 3mm disc sample of sapphire or } } magnesium orthovanadate. No glue, no mounting on a Cu ring, } } no nothing. } } } } ------------------------------------------------------- } } Laurence Marks } } Department of Materials Science and Engineering } } Northwestern University } } Tel: (847) 491-3996 Fax: (847) 491-7820 } } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } } ------------------------------------------------------- } } } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } } http://xraysweb.lbl.gov/esg/phasing/index.html } } } } } } } } } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html
-- Gordon Nord Small Business Network Design and Construction Macintosh and Windows - Solutions and Conflicts
Nord Consultants 20594 Cornstalk Terrace Ashburn VA 20147
Below is the result of your feedback form. It was submitted by (linda_knoll-at-msn.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 7, 2001 at 18:43:01 ---------------------------------------------------------------------------
Email: linda_knoll-at-msn.com Name: Linda Knoll
Organization: St.Joseph'sAcademy
Education: Graduate College
Location: St. Louis, MO USA
Question: How do the cells of humans and apes differ when looked at under a microscope?
Below is the result of your feedback form. It was submitted by (georgas1-at-home.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 7, 2001 at 20:29:09 ---------------------------------------------------------------------------
Email: georgas1-at-home.com Name: Adam Georgas
Organization: UMS-Wright Preparatory School
Education: 9-12th Grade High School
Location: Mobile, Alabama, USA
Question: Dear Microscopist: I am a 10th grade student working on my science fair project. My project deals with a computer program I wrote using fractal geometry to diagnose chronic lymphocytic leukemia. I obtained 25 pictures of normal cells taken from a Phillips electron microscope and 25 pictures of leukemic cells taken with the same microscope. I obtained these photographs from my mentor, a physician/professor at the University of South Alabama. My concern with my project is that the 25 healthy cells came from only one patient, and the 25 leukemic cells came from another patient. So, I have 50 cell pictures, 25 healthy, 25 leukemic, from only 2 patients. All of the research I have read use 25 different patients with only one cell from each patient. I am wondering if my study is valid? I posed this question to a local pathologist. She told me that since the pictures were taken from an electorn microscope, it was actually better to have more cells from just 2 patients. Is this true? If it is true, what is the rationale
Here is some parameters of 2000FX with AHP2OL polepiece: --------------------------------------------- Point resolution 0.31nm Lattice resolution 0.14nm OL focal length 4.1mm Cs 3.4mm Cc 3.1mm ---------------------------------------------
As to the question of beam semi-convergence angle, Prof. O'Keef has given a perfect note. I learnt a lot there as well.
Yours, Shu-You Li ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
Fax: +49-6131-3923768 Tel: +49-6131-3923148(O) E-mail: syli-at-mail.uni-mainz.de; syli16-at-hotmail.com URL: http://syli.homepage.com/ (This URL contains my resume, SCI journal informations, JobList, TEMAlert, as well as other useful informations related to Transmission Electron Microscopy) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Paula: Get in touch with Helmut Patzig at MOC, Spring Valley, NY. His email is Mocleica-at-aol.com. If anybody has the know-how and parts, I am pretty sure he does. Sorry I don't know of anyone closer to DC, but I have dealt with Helmut over the years with obsolete Reichert and LKB equipment.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Wed, 07 Mar 2001 08:31:14 -0500, Paula Sicurello 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 | -----------------------------------------------------------------------. | | | Hi Listers, | | I have a LKB Bromma 11800 Pyramitome whose belt has come loos and it needs to be replaced. I can't find the instruction manual or the slip of paper that my predecessor wrote the service person's name on. | | Does anybody out there have a source for the instruction manual? I called Leica but they told me this machine was absolete when they acquired the LKB line. | | I'll fix this thing myself if anyone can send me a copy of the manual (I'll glaly pay for the cost of copying and shipping). | | If anyone knows of a repair person who still works on these ancient beasties, please let me know. It would be good to have someone who could give them tune-ups every now & then. | | My screws are loose and I've got the hammer, blow torch sosme bubble gum and bailing wire....now all I need is the manual. | | | Pretty please?? | | Paula :-) | | Paula Sicurello | George Washington Univ. Medical Center | Dept. of Pathology, Ross Hall rm 505 | Electron Microscope Lab | 2300 Eye St. | Washington, DC 20037 | 202-994-2930 phone | 202-994-2518 fax | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
The answer depends on the details of the "TV" system and microscope used. At this point I don't even know if it is an optical or electron microscope, etc... It should be noted that actual microscope resolution and the displayed resolution via TV have little in common.
NTSC (US) TV is 525 lines vertical (total, and the equevelant of 280 to over 640 horizontal (bandwidth dependent). Note that not all 525 lines are used for the image.
PAL, a standard used in many other areas of the world is slightly higher vertical resolution, but not much.
There are other "non-brodcast standard" resolutions used by custom instrumentation.
Woody
} -------------------------------------------------------------- } ---------- } 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 } -------------------------------------------------------------- } ---------. } } } } } } } } } Email: de-at-payload.com } } Name: De McKeown } } } } Organization: Payload Systems } } } } Education: Graduate College } } } } Location: Cambridge, MA, USA } } } } Question: I need to determine the number of TV lines my } microscope is } } giving me. I have a test target but it only goes to 266 } l/mm and I have } } better resolution than that. I can't find a 'better' } target and am stuck } } trying to get an answer. How can I do this or where can I } go for a new } } target? } } } } I also have to find a contrast, but haven't got enough } information yet to } } ask a good question. } } } } Thank you!! } } } } }
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Tina, Check the following reference for your answer to your second question:
Liposits, Zs., D. Sherman, C. Phelix and W.K. Paull. (1986). A combined light and electron microscopic immunocytochemical method for the simultaneous localization of multiple tissue antigens. Histochemistry 85:95-106.
We very successfully used vibraotomed sections of brain to do light and EM ICC. Becuase of the penetration problems for the gold conjugates available at that time, we used PAP-DAB for visualization on the light level followed by silver intensification of the material for EM visualization. The study involved serial sectioning of the material to locate synapses so flat embedding of the sections was manditory. The method is fully discribed in the paper but feel free to contact me if you have questions.
I would also try using the ultra small gold colloids presently on the market followed by silver intensification as an alternative to the PAP-DAB technique.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
I have two questions for you immunolabeling experts.
First, a client has some precious and irreplaceable histological sections of human hippocampus that have been labeled with ABC - DAB. Will it be possible for me to de-paraffinize the sections, expose them to osmium vapor and re-embed them in resin on the slide and pop them off and resection them and see the DAB precipitate? Anyone have a protocol that has worked?
Second, these researchers plan to use Vibratome sections of mouse brain and immunolabel them for light microscopy using an ABC kit. I have done on-ultrathin-section colloidal gold immunolabeling for TEM. However, for this new project we would like to try pre-embedding labeling of 50-60 micrometer Vibratome sections, then embed and resection them for TEM. Knowing what works for light microscopy, we'd like to use streptavidin/colloidal gold or whatever for TEM visualization. My question is, of course, does anyone have a favorite protocol they would be willing to share? How do I keep the sections from curling? If we stick them on a glass slide to embed in resin and (hopefully) pop off later for resectioning, when and how do I get the to adhere?
Mahalo!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Michelle, The answer to the last question is easy. Remember Bugs Bunny munching his carrot? Rabbits eat plants so it is not too surprising that a preimmune serum recognizes a plant antigen, if not several.
What we have done in situations like this is to incubate the serum with the protein of interest (in this case, the 32kD glucanase) and then stain with that. This removes the specific glucanse staining. It is a kind of guilt by subtraction. It is not as nice as doing affinity purification, but it is a lot faster, and conserves the serum.
Hope this helps, Tobias Baskin
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Dear Listservers, we have been recently working on structural and compositional characterization of InGaAs/GaAs quantum dots grown by MOVPE, achieving interesting results. We have tried to use CERIUS2 to obtain simulations of cross sectional HREM images of these dots, but we experienced several problems. Is there anyone who has spent time on similar matters. We would like to discuss and compare our experiences, to understand if our problems are due to a bad construction of the supercell or to limitations of the software. Thanks Massimo
Thanks for all the responses to my original query. At the suggestion of Bart Cannon, you can check out my detector design at:
http://www.mta.ca/~jehrman/ted.htm
I perhaps should have clarified originally that this is a very simple design, and does not require any additional electronics, light pipes, PMT, etc. My machinist made it for me in an afternoon. I took inspiration from a single sentence in Goldstein, et. al (SEM and X-ray Microanalysis, 2nd edition, p. 267):
"A simple, inexpensive detector can be made from a high-atomic-number scattering surface placed below the specimen and tilted so that the transmitted electrons are scattered toward the conventional E-T detector in the specimen chamber"
} From my experience with one commercially available "real" TED a number of years ago, the homemade detector seems to perform as well (or better)
than the real thing.
No reference was given in the text, so I'm rather curious who came up with the idea originally. Are there any co-authors of the text lurking nearby who could shed any light?
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Question: Dear Microscopist: I am a 10th grade student working on my science fair project. My project deals with a computer program I wrote using fractal geometry to diagnose chronic lymphocytic leukemia. I obtained 25 pictures of normal cells taken from a Phillips electron microscope and 25 pictures of leukemic cells taken with the same microscope. I obtained these photographs from my mentor, a physician/professor at the University of South Alabama. My concern with my project is that the 25 healthy cells came from only one patient, and the 25 leukemic cells came from another patient. So, I have 50 cell pictures, 25 healthy, 25 leukemic, from only 2 patients. All of the research I have read use 25 different patients with only one cell from each patient. I am wondering if my study is valid? I posed this question to a local pathologist. She told me that since the pictures were taken from an electorn microscope, it was actually better to have more cells from just 2 patients. Is this true? If it is true, what is the rationale
Dear Adam, First, there is nothing about data from an electron microscope that makes sample selection any different from that used with other forms of data. The idea is that the population in your sample--in this case the normal and leukemic cells--should be representative of the larger population--all human normal lymphocytes and all human leukemic lymphocytes. If all normal lymphocytes in any one person have identical morphology, then any one cell will represent the population; if lymphocytes vary within an individual, then you would need a selection having the same distribution of types as exist in the whole person. This is usually obtained by selecting a small sample volume "at random". You might imagine how this could go wrong; e.g., if certain types were not present in arterial blood to the same extent as in venous blood. The next consideration is whether either normal lymphocytes or leukemic lymphocytes differ from person to person. I can see no reason that this would not be the case--especially if different forms of leukemia result from different transformation processes; e.g., different mutations or different extents of gene expression. In order to be sure that your sample is relevant, you would need to know whether there were different types of normal and leukemic cells both within and among individuals. This is an area where I am completely ignorant, but there is probably an extensive literature on this. There could also be differences arising from the preparation of the cells for EM, such as variation in fixation or staining, which can give apparent differences from nomonally identical cells. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Hi, I just started to bring up an ISI-SR50A. Followed the procedure to replace the filament, saturate beam current and aligment correctly but I could not get a decent image on the CRT even at low mag, 200x. Is there a way to check the detector? I varied the applied voltage from 5KV to 20KV and WD from 7mm to 40mm without any success. Any suggestions are welcome. -Vu.
__________________________________________________ Do You Yahoo!? Get email at your own domain with Yahoo! Mail. http://personal.mail.yahoo.com/
Here are two questions that I am pondering at the moment for thin film analysis in the TEM.
If you take the integrated intensities of the center portions of the peaks (zero crossing to zero crossing) of a top hat filtered spectrum to use for quantification, will you get the same results as you would if you do a background subtraction and take the integrated intensities from the peaks themselves?
For the integrated intensities of a top hat filtered spectrum, is it more appropriate to include the two negative lobes of the filtered peak added to the center portion?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
There are two freeware software packages that can accomplish this effort.
An IBM inspired software called OpenDX (for Unix), which is quite advanced. Start here (I have just installed this software but not worked with it yet):
http://www.opendx.org/
and through scion image, which is a PC version of NIH image. Start at the link below
http://rsb.info.nih.gov/nih-image/
You must install the ImageJ plugin, which you can obtain from:
http://www.isi.uu.nl/people/michael/vr.htm
This site also has other links for related information.
I am pursuing similar work with FIB/ Auger on particles and I would love to know about your progress. I believe this area has interesting and useful applications and I have found only very limited publications in this area. I would appreciate any additional information you might have.
Good Luck !
Regards, Ed
************************************************************ Edward Principe, Ph.D. Member of Technical Staff Defect & Thin Film Characterization Laboratory Applied Materials 408-986-3882
"Richard M Langford" {richard.langford-at-materials.oxford.ac.uk} on 03/08/2001 01:51:45 AM
To: microscopy-at-sparc5.microscopy.com cc: (bcc: Edward Principe/APPLIED MATERIALS)
Dear All
Can anyone suggest suitable software for 3-D reconstruction of grains and cracks from a sequential set of focused ion beam cross-sections.
Regards
Richard -------------------------------------------------------------- Richard M Langford
Department of Materials, University of Oxford Parks Road, Oxford, OX1 3PH, UK
If you only need to do reconstruction, download our free VoxBlast Light software at www.vaytek.com. Go to VoxBlast 3D software, then VoxBlast Light 3.0.
If you need any help or want more functionality, feel free to contact us directly.
Best regards,
Steve Niemela Sales sniemela-at-vaytek.com Ph: 641-472-2227 Fax: 641-472-8131
VayTek, Inc. 305 West Lowe Avenue Fairfield IA 52556 www.vaytek.com ----- Original Message ----- } From: "Richard M Langford" {richard.langford-at-materials.oxford.ac.uk} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 08, 2001 3:51 AM
A colleague is looking for a published reference that describes cholesterol crystals using transmission electron microscopy. We have searched the web but found only light microscopy of the crystals. Any help would be appreciated.
Thank you,
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 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 ##############################################################
Facility Scientist (Academic Assistant III) Flow Cytometry and Confocal Imaging Facility, Biotechnology Center
The Biotechnology Center and the Department of Molecular and Cell Biology at the University of Connecticut invite applications for the position of Facility Scientist for the Flow Cytometry and Confocal Imaging Facility. This Facility houses a Becton Dickson FACSCalibur Flow Cytometer/cell sorter, a Leica SP2 laser scanning confocal microscope and several other microscope and image processing workstations. The facility scientist will be responsible for working with University faculty and students to develop research projects that utilize these instruments and related technologies. The facility scientist will also organize periodic training sessions and workshops to familiarize new users with the instruments. This individual will also be encouraged to develop collaborative research projects with University faculty and with local biotechnology businesses. We seek an individual who is excited by the challenges of an interdisciplinary research group and who enjoys interacting with people in an active scientific environment. The preferred candidate will have a Ph.D. in the biological sciences and familiarity with one or both of the core technologies. This is a full-time, annually renewable position. Salary will be commensurate with experience and education. Submit curriculum vitae, a statement of experience and interests, and the names, addresses, telephone numbers and e-mail addresses of at least three references to: Biotechnology Center, Confocal Search Committee, University of Connecticut, 184 Auditorium Rd., Unit 3149, Storrs, CT 06269-3149. Applications may also be faxed to (860) 486-5005 or sent electronically to biotctr1-at-uconnvm.uconn.edu. Screening of applications will begin immediately and continue until the position is filled.
We encourage applications from under-represented groups including minorities, women and people with disabilities. (Search #01A372) --
************************************************************ Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
The following posting is for a colleague who is not on the listserver.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
We recently experienced a fire using a 5% perchloric acid - methanol electrolyte in a commercial jet electropolisher, and request information on similar experiences.
During operation of the electropolishing unit at room temperature and moderate pump speed, when turning up the voltage towards 30 V a loud pop was heard and the reservoir was found to be in flames. The fire was quickly extinguished using a dry fire extinguisher, but the reservoir and cable shielding melted.
We would appreciate any input on similar experiences using electropolishers with this or similar electrolytes. Any explanations for this behavior that you can provide would also be appreciated.
David S. Gelles Structural Materials Research Pacific Northwest National Laboratory Richland, WA 99353 Tel: (509) 376-3141 Fax: (509) 376-0418 E-mail: ds_gelles-at-pnl.gov
There are two ways that the top-hat filter has been used for quantifying EDS spectra:
(1) One can simply apply the filter to the spectrum as you suggest -- what results looks much like the second derivative of the original spectrum and each peak is represented by a central positive lobe with a negative lobe on either side. The slowly varying (continuum) components of the spectrum are suppressed. With care, one can relate the area of the positive lobe to the area of the original peak, however, there are some issues which make this simple concept a less than satisfactory quantitative method: (1) the area of the positive lobe is proportional to the area of the original peak, but the proportionality is a function of both peak area and filter shape. Thus the proportionality varies with the peak energy since the peak width varies with energy. (2) overlapping and adjacent peaks will interfere with each other, and this interference is actually increased by the broadening effect of the filter. Oddly enough, I have seen a number of familiar reference works which seem to give me credit for inventing the "top hat filter" used in this way -- I didn't. I saw it used this way for nuclear gamma ray spectra in the early '70s and after trying it myself, realized that it really wasn't satisfactory. But this experience did lead to the "filter-fit" method which follows.
(2) The"right" way for using the top-hat filter for continuum suppression is to use it in conjunction with linear-least squares fitting. One simply filters the unknown and each of the reference spectra to be fitted to it and then applies a conventional linear fit between them. When one fits the filtered references to the filtered unknown, the fact that the spectra have been distorted by the filter is immaterial (since the filter operator is linear), the overlaps are accurately unfolded, and the continuum basically drops out of the problem. Properly implemented, this is a very good quantitiative technique and has been employed productively for almost 30 years. If interested, the technique is published in NBS 604 (P273) or interested parties could contact me for references to several other papers describing it.
Fred Schamber
"Walck, Scott D." 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 } -----------------------------------------------------------------------. } } Here are two questions that I am pondering at the moment for thin film analysis in the TEM. } } If you take the integrated intensities of the center portions of the peaks (zero crossing to zero crossing) of a top hat filtered spectrum to use for quantification, will you get the same results as you would if you do a background subtraction and take the integrated intensities from the peaks themselves? } } For the integrated intensities of a top hat filtered spectrum, is it more appropriate to include the two negative lobes of the filtered peak added to the center portion? } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } 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)
I've been gold immunolabelling whole retina pieces - 300um thick sheets - with great success (LM and EM) for years. I react the tissue in small vials, floating around loose. Briefly, you need a fixative like PLP or paraformaldehyde - any glut in the fix and antibody access to the tissue is decreased and you just get labelling on the cut edges. Use saponin in all solutions for penetration enhancement. Use an Fab2 anti Ig conjugated to 1 nm gold. Silver enhance then gold tone. Pop tissue in a silicon mould for embedding. If this sounds useful I can send you the full method.
As for rescuing paraffin sections, I did some a long time ago. From memory the resulting tissue looked like squashed newspaper - a mushy black mess with a hint of structure. I can't imagine even being able to distinguish the DAB from the rest, let alone see what was labelling. However, maybe methods have improved.
Cheers,
Diana --
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
In my opinion what happened was that a spark between the cathode and specimen, which often occurs at those voltages during electropolishing, ignited the electrolyte. I have used other electrolytes at voltages greater than 30V and I have also head a crackling noise which indicated to me that was sparking. Fortunately the electrolyte was not as ignitable as a 5% perchloric acid - methanol electrolyte. Nevertheless, I kept the voltage below that just in case for the electrolyte was still based on organic solvents.
You are going to be bombarded by horror stories and cautions about electrolytes containing perchloric acid. And they are right. I will not say that I have used electrolytes containing up to 30 vol.% in ethanol without problems because that would be tempting fate and I have enough Sicilian blood in me to make me a bit superstitious.
I have some recommendations.
1. As far as I know, perchloric acid electrolytes only require a voltage of 15V max. It is rare that I have used voltages in excess of 25V for any electrolyte and that was with an electrolyte of methanol, Butoxy-ethanol, magnesium perchlorate and lithium chloride (LiCl). This was the electrolyte where above 30V I was getting sparking. I have used water based electrolytes above 30V sometimes, but ignition is not an issue there.
2. Whenever making up or using an electrolyte containing perchloric acid, cool it down. When making it up, this prevents ignition due to heat of dissolution of the concentrate acid as it mixes. This may be paranoia. Cooling during electropolishing is necessary because it also reduces the risk if ignition and it improves the electropolishing process. I have found that most organic based electrolytes, especially those based on ethanol and methanol require cooling to {20°C in order to achieve the correct electropolishing conditions. So the necessity of cooling is two fold.
I hope this is useful.
"Thomas, Larry (PNNL)" 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 } -----------------------------------------------------------------------. } } The following posting is for a colleague who is not on the listserver. } } Larry Thomas } Pacific Northwest National Laboratory } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto:Larry.Thomas-at-pnl.gov } } We recently experienced a fire using a 5% perchloric acid - methanol electrolyte } in a commercial jet electropolisher, and request information on similar } experiences. } } During operation of the electropolishing unit at room temperature and moderate } pump speed, when turning up the voltage towards 30 V a loud pop was heard and } the reservoir was found to be in flames. The fire was quickly extinguished } using a dry fire extinguisher, but the reservoir and cable shielding melted. } } We would appreciate any input on similar experiences using electropolishers with } this or similar electrolytes. Any explanations for this behavior that you can } provide would also be appreciated. } } David S. Gelles } Structural Materials Research } Pacific Northwest National Laboratory } Richland, WA 99353 } Tel: (509) 376-3141 } Fax: (509) 376-0418 } E-mail: ds_gelles-at-pnl.gov
-- Dr. Steven Celotto Department of Engineering Materials Science & Engineering University of Liverpool Brownlow Hill Liverpool L69 3BX UNITED KINGDOM
I don´t know about the ISI50A but, assuming that it has the typical Everhard-Thornley design, isn´t it just possible that the bias voltage is inverted in your detector?. If the bias voltage is set negative, it´ll be rejecting all the secondary electrons, and you would only get a generally noisy image composed of the primary electrons backscattered in the very direction of the detector´s position. Quite nice for topographic information about flat specimens, but far worse quality than in "normal" SE images. Pardon me if I raised a too obvious explanation, but it occurred to me sometimes to get puzzled with a bad functioning of my SE detector just because another user of the microscope had been playing with the knobs.
Cheers
Diego
Diego Alvarez Instituto Nacional del Carbón INCAR-CSIC Spain
Thanks to all who responded to may plea for help in finding either a manual or a service person for the wee beastie.
Slowly but surely I will tighten the screws.....
Thanks again!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Full details of Trends in Nanotechnology (TNT) 2001, Segovia, Spain, September 2001, are now available. Based on the same format as last year, the conference aims to bring together researchers from all disciplines relating to nanotechnology, in a setting with ample time for interaction. A new feature this year will be the availability of grants to enable graduate students to attend and present posters.
Full details can be found at www.cmp-cientifica.com/tnt2001
Regards
Tim
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
The NIST's web link to DTSA can be found at: http://www.cstl.nist.gov/div837/837.02/MicroscopySoftware.html
This program is free and available for the Macintosh only.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-417-1321 100 Bureau Drive, Stop 8371 Gaithersburg, MD 20899-8371 http://www.nist.gov/cstl/div837/837.02/
TO be more specific, any advice on using formvar coated slot grids for serial sections. They seem to crinkle, or get blobby, right on the vessel I want to look at.
Ellen: Another question: are you using bare formvar or have you coated with carbon? Bare formvar will pucker, but the addition of a little carbon does wonders for stabilization and strength of the films. My experience is using 0.5% formvar, stabilized with about 10nm of evaporated carbon. These films aren't too thick, allowing good image resolution at 80kV. BTW: if the formvar films are puckered, they will show it prior to picking up sections. (Another vagrant neuron just fired.) One way to deal with the puckered formvar is to use chloroform or carbon tet. Simply pass the grid over the mouth of an open bottle, or, as a last resort, put the grid into the neck of the bottle (NOT into the fluid--just the vapors). Please be careful with this. (Exposure to these chemicals.) I may have the beginnings of interstitial pulmonary fibrosis--a not uncommon result of exposure to chemicals.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
On Sun, 11 Mar 2001 14:31:50 -0500, Ellen S. Morgan 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 | -----------------------------------------------------------------------. | | | TO be more specific, any advice on using formvar coated slot grids for | serial sections. They seem to crinkle, or get blobby, right on the vessel | I want to look at. |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Does anyone know of any buildings/laboratories recently constructed that house instruments (not necessarily EM's) requiring very low ( {0.5 mG) 60 Hz stray magnetic fields? I am interested in contacting someone (scientist, building manager, architect, etc.) who might know the details of how that building was constructed.
Thank you for your time and help,
Sincerely,
Mick Thomas ------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
At 5:24 PM +0100 3/9/01, Edmund Gierlik wrote: } Email: gierlik-at-delta.sggw.waw.pl } Name: Edmund Gierlik } Organization: Electron Microscopy Lab. } Education: professor } Location: Warsaw, Poland } } Question: Can anybody show the source of nice presentation of ESEM } application in a phase transition investigation (water - ice, liquid - solid)?
Trisha Rice at the ESEM applications lab might be able to provide this. She can be reached at either trice-at-feico.com or trice-at-electroscan.com (I'm not sure if this last address is still valid).
Dear Randy, I have just a few points to add to Steve's excellent advice.
1. "It is too dim to focus" - then i) increase the emission current, many people run at too low a current and make the task of optimising the instrument settings much more difficult ii) increase the kV which would increase the intensity too ii) re calibrate the photographic system to allow a focus intensity that is suitable for most operators
I often use LoDose film, which is more than an order of magnitude more sensitive than SO163, so I can barely see the beam on the screen. Fortunately, we have an intensified CCD, which is so sensitive that I can still focus. In fact, using the minimum-contrast method works very well with this system, since the contrast can be electronically enhanced.
6. To focus at low magnifications try removing the objective aperture and focus without a binocular, or a wobbler, looking for the very strong minimum contrast effect.
Furthermore, I discovered that inserting the diffraction aperture (with the objective aperture out) allows this method to be used at up to 63kx (and possibly higher). Since minimum contrast can easily be found within one second, this is well suited to radiolabile specimens.
Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
This is a different procedure from the Formvar-coated grids that you are using but it's one of my favorite journal articles/techniques - J. C. Rowley and D.T. Moran, A Simple Procedure for Mounting Wrinkle-Free Sections on Formvar-Coated Slot Grids. Ultramicrotomy 1 (1975) 151-155. They use Formvar-coated aluminum supporting racks. We use this method for slot grids all the time rather than Formvar-coated grids. good luck, Beth } TO be more specific, any advice on using formvar coated slot grids for } serial sections. They seem to crinkle, or get blobby, right on the vessel } I want to look at.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I do quite a bit of serial sectioning and would like to offer you the following advice:
1) Cut your sections as small as possible, they can be as much as 1mm wide, but try to make them something like 0.1mm high. This will allow you to get you as many as 20 sections on a 2mm slotted grid.
2) Get a bunch of locking tweezers and from the time you pick up the grid until you put it in the microscope, do not put the grid down onto filter paper (as you would for a normal grid). So, after you pick up the ribbon of sections, leave the grid in the tweezers to dry, and then when you post stain the grids, leave them in the tweezers to dry. If you put them down onto filter paper the formvar may rupture.
3) Maintain a positive attitude, and if things aren't going your way, stop and come back to it another day (the blocks will always be there for you, but if you get burned out you won't be there for the blocks).
Best of Luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
NATIONAL INSTITUTES OF HEALTH Bethesda, Maryland Division of Bioengineering & Physical Science Supramolecular Structure & Function Lab.
Biologist GS9/GS11/GS12 trained in life sciences and/or physical sciences at BS or MS level with solid technical experience in thin-section transmission electron microscopy. Experience in advanced techniques in analytical electron microscopy and structural biology (cryosectioning, high-pressure freezing, and macromolecular preparation methods) is desirable, although on-the-job training is possible for an experienced microscopist. PROOF OF U.S. CITIZENSHIP IS REQUIRED for this appointment.
For further information please contact:
Dr. Richard Leapman Division of Bioengineering & Physical Science Bldg. 13, Rm. 3N17 National Institutes of Health Bethesda, MD 20892 Tel: (301) 496-2599 FAX: (301) 496-6608 E-mail: leapman-at-helix.nih.gov
Applications (with curriculum vitae or federal employment form SF-171) should include the reference number ORS-01-0044, and should be post marked by April 9, 2001 and sent to:
Attention: Mr. Harold Atkins National Institutes of Health ORS Human Resources Office 31 Center Drive Msc 2157 Bldg 31, Room 4b41 Bethesda MD 20892-2157
Many thanks to everyone who responded to my question about focusing. Although my actual question was answered almost immediately, the ensuing replies were very enlightening. In EM work, it seems, even the "simplest" things have dimensions that aren't always obvious.
My initial question arose from the fact that I was curious about something I had always done as a matter of course because I had been taught to do it that way. That is, I have spent years focusing TEM's with the beam at the crossover point because I believed that I'd been taught to do so because of some property of electron "optics" that made it the preferred method to ensure best focus. Turns out, though, that the reason was something else entirely, involving an earlier generation of microscopes that simply had a beam that was too dim for focusing at the intensity needed for recording an image.
It's always educational to question old assumptions. Embarrassing, too, sometimes....
Thanks again, listers.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Technically, re-embedding paraffin sections for EM is perfectly feasible. The way I have done it is to separate the sections from slides after re-hydration and before further fixation with glutaraldehyde and osmium. Since both glutaraldehyde and osmium treatment cause sections to shrink and be brittle, sections might get further torn if they are still attached to the slides. Another step you should be careful about is to keep sections flat when applying glutaraldehyde and osmium. This ensures sections not to curl or wrinkle later. You can float your sections in 0.1 M phosphate buffer in a Petri dish, and let sections lie flat on the bottom. Then you gently remove buffer and add fixative with a glass pipet without disturbing the sections. Do not shake sections for a while after adding fixative. To flat-embed the sections, you can sandwich sections that have been fully infiltrated with resin between two sheets of Aclar film, then place this sandwich between two glass slides and apply some weight on top while polymerizing the resin. As someone already pointed out, even though you can re-embed paraffin sections for EM, ultrastructural will not be adequate anymore. This is particularly a problem for examining brain tissue in which membrane outline is essential for identifying different neuronal elements. However, if the tissue is precious, I would say it is still worth trying. We work with post-mortem brain tissue often. Sometimes the brains were over twenty-four hours post-mortem, fixed with only formalin and stored in cryoprotectant in a freezer for a long time. I am often surprised how much information we can still obtain from such tissue at EM level. I would be happy to help you to evaluate your micrographs if you like.
Good luck, Tina, let me know if you need more detailed answers.
Hong
============== Hong Yi Supervisor Emory University School of Medicine Neurology Microscopy Core Laboratory Rm 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
---------- } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: TEM - immunolabeling } Date: Wed, Mar 7, 2001, 2:55 PM }
} ------------------------------------------------------------------------ } 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 two questions for you immunolabeling experts. } } First, a client has some precious and irreplaceable histological sections } of human hippocampus that have been labeled with ABC - DAB. Will it be } possible for me to de-paraffinize the sections, expose them to osmium } vapor and re-embed them in resin on the slide and pop them off and } resection them and see the DAB precipitate? Anyone have a protocol that } has worked? } } Second, these researchers plan to use Vibratome sections of mouse brain } and immunolabel them for light microscopy using an ABC kit. I have done } on-ultrathin-section colloidal gold immunolabeling for TEM. However, for } this new project we would like to try pre-embedding labeling of 50-60 } micrometer Vibratome sections, then embed and resection them for } TEM. Knowing what works for light microscopy, we'd like to use } streptavidin/colloidal gold or whatever for TEM visualization. My question } is, of course, does anyone have a favorite protocol they would be willing } to share? How do I keep the sections from curling? If we stick them on a } glass slide to embed in resin and (hopefully) pop off later for } resectioning, when and how do I get the to adhere? } } Mahalo! } } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
Regarding your question about pre-embedding immunolabeling of vibratomed brain sections, the type of labeling to use depends on the questions the researchers are trying to answer. Immunoperoxidase gives greater labeling depth, but limited spatial resolution. If they simply want to know what neuronal elements are labeled, then immunoperoxidase is fine. Otherwise, imunogold should be considered because it provides much more precise localization.
The methods for pre-embedding immunogold labeling using vibratomed brain sections have been very well documented. One of the first publications was by Chan, Aoki and Pickel (1) at Cornell University in 1990, shortly after ultrasmall gold conjugates were introduced. Many brain researchers have adapted their protocol. The characteristics of this protocol are the use of acrolein in the fixative for perfusion, using little or no detergent treatment for permeabilization and using a 1 nm colloidal gold conjugated IgG secondary probe in conjunction with a light insensitive, low viscosity, but fast reacting silver enhancement reagent. I have seen many beautiful results using this protocol. However, the particle size tends to be big, and irregular in shape. The cause of this is mainly in the silver enhancement reagent. In 1999, a new EM grade silver enhancement reagent was introduced to the market. Because of the improved enhancement homogeneity in particle size and shape, this silver enhancement reagent allowed us to conduct for the first time the pre-embedding double immunogold labeling in brain tissue using only ultrasmall gold conjugates (2).
To avoid a lengthy posting, I will send you the protocol, along with images off-line.
1. Chan J. Aoki C. Pickel VM. (1990) Optimization of differential immunogold-silver and peroxidase labeling with maintenance of ultrastructure in brain sections before plastic embedding. Journal of Neuroscience Methods. 33(2-3):113-27,
2. Hong Yi, Jan L.M. Leunissen, Ge-Ming Shi, Claire-Anne Gutekunst, and Steven M. Hersch (2001) A Novel Procedure for Pre-embedding Double ImmunogoldSilver Labeling at the Ultrastructural Level J. Histochem. Cytochem. 2001 49: 279-284.
Hong
============== Hong Yi Supervisor Emory University School of Medicine Neurology Microscopy Core Laboratory Rm 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
---------- } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: TEM - immunolabeling } Date: Wed, Mar 7, 2001, 2:55 PM }
} ------------------------------------------------------------------------ } 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 two questions for you immunolabeling experts. } } First, a client has some precious and irreplaceable histological sections } of human hippocampus that have been labeled with ABC - DAB. Will it be } possible for me to de-paraffinize the sections, expose them to osmium } vapor and re-embed them in resin on the slide and pop them off and } resection them and see the DAB precipitate? Anyone have a protocol that } has worked? } } Second, these researchers plan to use Vibratome sections of mouse brain } and immunolabel them for light microscopy using an ABC kit. I have done } on-ultrathin-section colloidal gold immunolabeling for TEM. However, for } this new project we would like to try pre-embedding labeling of 50-60 } micrometer Vibratome sections, then embed and resection them for } TEM. Knowing what works for light microscopy, we'd like to use } streptavidin/colloidal gold or whatever for TEM visualization. My question } is, of course, does anyone have a favorite protocol they would be willing } to share? How do I keep the sections from curling? If we stick them on a } glass slide to embed in resin and (hopefully) pop off later for } resectioning, when and how do I get the to adhere? } } Mahalo! } } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
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A user has encountered an interesting problem . When cells are scraped and pelleted and embedded directly in epoxy they appear OK under the scope. When they are enrobed in agarose and then embedded the appearance becomes variable, although most cells appear very dense with contrast between organelles very low. This variability in appearance can be on the same grid square. Any ideas? Thanks Bruce Bruce Cutler Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id NAA02400 for dist-Microscopy; Mon, 12 Mar 2001 13:48:03 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id NAA02392 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 12 Mar 2001 13:47:33 -0600 (CST) Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu [134.193.71.1]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id NAA02384 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 12 Mar 2001 13:47:22 -0600 (CST) Received: by email.exchange.umkc.edu with Internet Mail Service (5.5.2653.19) id {FCF6B0PG} ; Mon, 12 Mar 2001 13:45:30 -0600 Message-ID: {95A711A70065D111B58C00609451555C01CA7611-at-UMKC-MAIL02} "'Microscopy-at-sparc5.microscopy.com '" {Microscopy-at-sparc5.microscopy.com}
May be it's not really a phase transition, but you can find pictures of dissolution and crystallization of table salt at our web page: http://www.umkc.edu/dentistry/microscopy/Esemimg.htm
Vladimir
-----Original Message----- } From: Edmund Gierlik To: Microscopy-at-sparc5.microscopy.com Sent: 3/9/01 10:24 AM
Email: gierlik-at-delta.sggw.waw.pl Name: Edmund Gierlik
Organization: Electron Microscopy Lab.
Education: professor
Location: Warsaw, Poland
Question: Can anybody show the source of nice presentation of ESEM application in a phase transition investigation (water - ice, liquid - solid)?
Dear listers, I have a customer who wants TEM on a section of cellulose that is coated with a layer of Fe2. I have no idea about how to go about accomplishing this. He wants to see where the Fe2 is located in the cellulose matrix, and how deep it has penetrated. The sample is a layer of cellulose about 1/2 mm thick with a visible layer of Fe on the surface and it's in submerged in water. Any ideas out there? Thanks so much,
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
After several requests, I've posted a measured schematic of my transmitted electron adapter, which can be reached via a link from the page mentioned previously:
http://www.mta.ca/~jehrman/ted.htm
I had the adapter made of aluminum and brass (just because the machinist likes to work with these). The angled surface below the specimen grid is polished brass, and as mentioned by several respondents, could be plated/coated/covered with something that generates more secondaries. Signal, however, has not been a problem in our applications - more than enough even at the smallest spot sizes.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I have a LKB Ultrotome V which has given many years of faithful service to me and my students until today, when the band that raises and lowers the cutting arm snapped. I am in the middle of the semester class on EM techniques and hence am a bit desperate to get it repaired. Does anyone know where I might find a replacement part for this - and is it possible to replace and adjust it myself? There are no instructions in my manual for this procedure; are there any out there somewhere? Is there any company that will service this instrument? I am located in New England.
Many thanks in advance for any leads.
Sincerely,
Dick Briggs Biology Department Smith College Northampton, MA 01063
This notice is to invite you to attend and participate in the 28th Annual Meeting of the Microscopical Society of Canada. This three day meeting will be held at the University of New Brunswick, Fredericton, New Brunswick, Canada on 6-8 June, 2001. The Scientific Program features invited speakers from the environmental sciences, plant sciences, material sciences, EELS applications, confocal microscopy and magnetic resonance imaging (MRI). Workshops will focus on specimen preparation techniques for both the biological and material sciences, including ultramicrotomy, polishing methods, high-pressure freezing techniques and a workshop on preparation of digital images for publication. Vendors and manufacturers will be demonstrating the latest developments in the field of microscopy. Presentations can be in either platform or poster format and the Abstract Deadline is March 30, 2001. The deadline for Pre-registration is April 30, 2001. All information and required forms for registration and submission of abstracts are available at the conference website at: http://www.unb.ca/msc2001. ************ Keynote Speakers and topics include: Dr. Bruce Balcom, Department of Physics, University of New Brunswick. Title of talk: "Magnetic Resonance Imaging of Materials". Dr. Ron Gronsky, Materials Science and Engineering, University of California, Berkeley. Title of talk: TBA. Dr. Michael Hochella, Dept. of Geological Sciences, Virginia Polytechnic Institute & State University, Blacksburg, Virginia. Title of talk: "Contributions of Microscopy to the Environmental Sciences: From Bacteria to Atoms" Dr. Richard Howard, DuPont Agricultural Products, Experimental Station, Wilmington, Delaware. Title of talk: "Trends in imaging fungal pathogens for understanding the cell biology of plant disease" Dr. Peter Ottensmeyer, Medical Biophysics, University of Toronto. Title of talk: "3-D Reconstruction of Macromolecules from single-particle STEM images: Transmembrane signalling of the insulin receptor"
Invited Speakers include: Mr. Jason Davis, Medical Biophysics, University of Toronto. Title of Talk: "EM of Chromophores: Very Low Energy-Loss Imaging of Green Fluorescent Protein and other Coloured Denizens." Dr. Gianluigi Botton, Materials Technology Laboratory, Natural Resources Canada. Title of Talk: "Analytical TEM of Macrostructures and Nanostructures". Dr. Elaine Humphrey, Biosciences EM Facility, University of British Columbia. Topic: "Confocal Microscopy" Dr. John McCaffrey, Institute for Microstructural Sciences, National Research Council Canada. Title of talk: "Structural Characterisation of InAs/GaAs and InAs/InP Quantum Dots by TEM" Dr. Doug Perovic, Dept. of Metallurgy & Materials Science, University of Toronto. Topic: "EM of Semiconductors". ************ Fredericton, New Brunswick is a three hour drive north from Bangor, Maine and an eight hour drive from Boston or Montreal. June is a beautiful time in New Brunswick and you can find interesting things to do and see through the conference website tourism links.
We turned water to ice on plant surfaces using the Peltier stage. We did not take any nice pictures (I have a few ugly ones!). It was an idea re ice nucleation on plants for a student project idea that we did not follow up.
Dave
On Mon, 12 Mar 2001 13:45:29 -0600 "Dusevich, Vladimir" {DusevichV-at-umkc.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 } -----------------------------------------------------------------------. } } } May be it's not really a phase transition, but you can } find pictures of dissolution and crystallization of } table salt at our web page: } http://www.umkc.edu/dentistry/microscopy/Esemimg.htm } } Vladimir } } -----Original Message----- } } From: Edmund Gierlik } To: Microscopy-at-sparc5.microscopy.com } Sent: 3/9/01 10:24 AM } Subject: ESEM-Need presentation of phase transition. } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } } Email: gierlik-at-delta.sggw.waw.pl } Name: Edmund Gierlik } } Organization: Electron Microscopy Lab. } } Education: professor } } Location: Warsaw, Poland } } Question: Can anybody show the source of nice presentation of ESEM } application in } a phase transition investigation (water - ice, liquid - solid)? }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I need to fix live cells (mainly blood cells) on a microscope glass slide to prevent them from movement but to maintain their viability for 1-3 hrs. Also it should be non-stained with any dye cells in mono-layer. Material to fix cells should be optically transparent in 500-700 nm. Room temperature is asssumed.
Any input about applicable technologies (gels etc) with references about suppliers of materials that are involved and sources for description of those techniques will be highly appreciated.
Thanks in advance
Dmitri Lapotko, Ph.D.
Luikov Heat and Mass Transfer Institute 15, Brovka Street Minsk, 220072 Belarus
----- Original Message ----- } From: "Patton, David" {David.Patton-at-uwe.ac.uk} To: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} Cc: "'Edmund Gierlik '" {gierlik-at-delta.sggw.waw.pl} ; {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, March 13, 2001 4:07 AM
} I am getting confused about light sources and Ca ratio imaging. I } have talked to people who use standard Hg arc sources and seem to } have no problems. Others seem to feel that you need a stabilized } power supply or a Xenon source for stability (see below). Are we } talking about line voltage variations (which a line } conditioner/voltage regulator ought to deal with) or actual arc } source variation? Any opinions out there? Thanks- Dave
} From my observations of using both types of light source, the Xenon seems } far more stable with time. You don't notice the fluctuations that you get } with Hg bulbs. This is important for quantification, such as dual excitation } ratio methods (eg. Fura-2). } } Stephen H. Cody, } Colon Molecular and Cell Biology Laboratory, } Ludwig Institute for Cancer Research, } Post Office Royal Melbourne Hospital, } Parkville, Victoria 3050, Australia. } } Tel: 61 3 9341 3155 Fax: 61 3 9341 3104 } email: stephen.cody-at-ludwig.edu.au } www.ludwig.edu.au/confocal } } } } ---------- } } From: Tom Phillips[SMTP:PhillipsT-at-MISSOURI.EDU] } } Reply To: Confocal Microscopy List } } Sent: Wednesday, 28 February 2001 2:08 } } To: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU } } Subject: Re: Xenon lamps } } } } All true but I believe one disadvantage is the peak illumination is } } less for some fluorochromes such as DAPI. We see a difference in our } } two systems that have Hg or Xe. } } } }
--
************************************************************ Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Greetings, Two standard points. These are not about power output but do relate to the choice of arc type (archtype??) which seemed to be on the questioner's mind. Forgive me if this is too obvious.
One is that the output of a xenon arc is much more uniform across the spectrum compared to the steep peaks of the mercury arc. For ratios with different excitation wavelengths, in principle difference in intensity between the two wavelengths doesn't matter, but in practice when the intensities differ alot, there can be problems. Therefore, if you are going to be working with a variety of wavelength pairs (lots of different dyes and applications) the continuity of the xenon will probably be helpful; whereas, if you will stick to a single pair of wavelengths, or ratio emitted light, then even spectral quality might not matter.
The other issue is that xeon arcs tend to give off lots more ozone than mercury arcs and so usually require significant venting. This can be a nuisance depdending on the lay of the land, etc.
As ever, Tobias Baskin
} } } I am getting confused about light sources and Ca ratio imaging. I } } have talked to people who use standard Hg arc sources and seem to } } have no problems. Others seem to feel that you need a stabilized } } power supply or a Xenon source for stability (see below). Are we } } talking about line voltage variations (which a line } } conditioner/voltage regulator ought to deal with) or actual arc } } source variation? Any opinions out there? Thanks- Dave } } } } } From my observations of using both types of light source, the Xenon seems } } far more stable with time. You don't notice the fluctuations that you get } } with Hg bulbs. This is important for quantification, such as dual excitation } } ratio methods (eg. Fura-2). } } } } Stephen H. Cody, } } Colon Molecular and Cell Biology Laboratory, } } Ludwig Institute for Cancer Research, } } Post Office Royal Melbourne Hospital, } } Parkville, Victoria 3050, Australia. } } } } Tel: 61 3 9341 3155 Fax: 61 3 9341 3104 } } email: stephen.cody-at-ludwig.edu.au } } www.ludwig.edu.au/confocal } } } } } ---------- } } } From: Tom Phillips[SMTP:PhillipsT-at-MISSOURI.EDU] } } } Reply To: Confocal Microscopy List } } } Sent: Wednesday, 28 February 2001 2:08 } } } To: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU } } } Subject: Re: Xenon lamps } } } } } } All true but I believe one disadvantage is the peak illumination is } } } less for some fluorochromes such as DAPI. We see a difference in our } } } two systems that have Hg or Xe. } } } } } } } } -- } } ************************************************************ } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 75 N. Eagleville Rd. U-125 } Storrs, CT 06269-3125 } Knecht-at-uconnvm.uconn.edu } 860-486-2200 860-486-4331 (fax) } home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html } ************************************************************
It could be a problem with penetration through the agar if cells are fixed in glutaraldehyde after the agar is added. Glut crosslinks the agar so that other solutions, including osmium and washes, can't penetrate properly. (I know this from experience.) We rinse the monolayer briefly and gently with warmed buffer (pour on, swirl, pour off). The buffer should be 300 mOsm (check with an osmometer) and pH 7.2-7.4 (check with a pH meter). Pour on the glut in buffer and let sit about 3 min (no more than 5); scrape, pellet cells in the glut and let sit for another 30 min. The pellets should be small (2 mm or so). If pellets are large, you can loosen them with the applicator stick so that the glut can get around to the underneath side. Just try not to disturb the clump too much. Remove glut with pipette. Scrape out pellet with flattened end of applicator stick. Enrobe with agar and then wash and stain as you would a piece of tissue. If there's a lot of excess agar, you can trim it away with a razor blade. This keeps them together and makes life much easier.
Some folks suspend cells in warm agar or gelatin before fixation and spin them down in the agar or gelatin and then fix. We have had variable staining this way, particularly if there is a lot of excess agar or the agar solidifies before the cells get to the bottom of the tube. If you can get a tight pellet by centrifuging fast in a warmed centrifuge (or jacketed tube) before the agar solidifies so that the layer of agar around each cell is thin, it will work. Do cut away the excess agar at the top of the tube, and make cell blocks small.
We centrifuge in a microfuge tube that looks like this:
| | | | | | | | | | \ / | | | | U
(Sorry about the graphics.) I think they come from PGC Scientific (800 424 3300). Sorry, I don't have the catelog number; our bag has been emptied, and our catelog has been "borrowed." They are about 2 " long and 3 mm in dia; the inside of the very tip is the diameter of a paper clip. This info was provided a while back, and maybe you can find it in the archives.
Centrifuge at right angles so that the pellet doesn't get stuck on the slanted neck of the tube. Then cut off the very bottom (below the cells) and then cut off the tube above the cells. Push out the cell "log" with a straightened paper clip. Chop into smaller logs.
Good luck. Address below if further questions.
On 12 Mar 2001, Bruce Cutler wrote:
} Date: 12 Mar 2001 12:58:48 -0600 } From: Bruce Cutler {bcutler-at-eureka.idl.ukans.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Agarose enrobed 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 } -----------------------------------------------------------------------. } } } A user has encountered an interesting problem . When cells are scraped and } pelleted and embedded directly in epoxy they appear OK under the scope. When } they are enrobed in agarose and then embedded the appearance becomes variable, } although most cells appear very dense with contrast between organelles very low. } This variability in appearance can be on the same grid square. } Any ideas? } Thanks } Bruce } Bruce Cutler } Director, Microscopy & Electronic Imaging Laboratory } University of Kansas } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Dear Mary Gail, Do you mean a layer of metallic iron? I think you should treat the cellulose as wood or plant tissue and fix and embed, then cut a cross-section as you would a plant stem. No staining necessary. You need a TEM or STEM with an EDX on it and it will easily locate the iron to the ten nanometer scale. At 03:05 PM 3/12/01 -0500, you wrote: } } Dear listers, } I have a customer who wants TEM on a section of cellulose that is coated } with a layer of Fe2. I have no idea about how to go about accomplishing } this. He wants to see where the Fe2 is located in the cellulose matrix, } and how deep it has penetrated. The sample is a layer of cellulose about } 1/2 mm thick with a visible layer of Fe on the surface and it's in } submerged in water. } Any ideas out there? } Thanks so much, } } Mary Gail Engle
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 e-mail: mager-at-interchg.ubc.ca
Hello, I was looking at some piping in our lab while sitting on the windowsill and noticed a sign painted over. Looking closer it was a sign for asbestos insulation that has been 80% painted over. I was told earlier that there was no asbestos pipe insulation in the lab. I collected some dust samples from near the fraying pipe insulation, on top of shelves, floor, and the windowsill and imaged them in our SEM.
Can anyone with some experience with asbestos take a look at the images at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the images looks like a candidate for asbestos? I think only fd6, fd7, and fd8 are asbestos, and the rest of the fibrous materials imaged are cellulose binding agents used in the insulation.
I will examine the material by TEM and electron diffraction, but I was wondering if someone could post the techniques they use for sampling from the dust, and any other hints and techniques I should be aware of.
Thanks for your help, Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
We have some data from a Wyko optical profilometer which we would like to work with offline & import into MATLAB for some further analysis. The lab tells me that the Wyko data can be output in "OPD" format. Does anyone know how we can covert this to a format we can work with.
Thanks
Tim
------------------------------------------------------------------------- Tim E. Harper CEO CMP Cientifica Tel. +34 91 640 7185 Fax. +34 91 640 7186
Gordon's comments concern me on several levels. The SEM, especially without EDX, is not the best tool to identify asbestos fibers. The TEM and diffraction can be absolutely definitive. However, I'm unaware of any EPA or NIOSH protocol for taking dust to the TEM grid, though one could be developed easily enough for Gordon's purposes. Gordon described friable pipe insulation with an asbestos warning. Seems wiser to bring his administration in on this situation. Does the UC system have some in-house personnel that can run a PLM for him? The lack of such in house personnel or an office dealing with asbestos abatement would be surprising for such a large university. Even a commercial lab PLM is faily inexpensive. While the amount of dust given off the pipe, barring some regular banging on the pipe, is probably minimal, the wiser course of action is bringing in someone who is well trained in dealing with asbestos pipe insulation. It doesn't have to be removed, just managed.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, I was looking at some piping in our lab while sitting on the windowsill and noticed a sign painted over. Looking closer it was a sign for asbestos insulation that has been 80% painted over. I was told earlier that there was no asbestos pipe insulation in the lab. I collected some dust samples from near the fraying pipe insulation, on top of shelves, floor, and the windowsill and imaged them in our SEM.
Can anyone with some experience with asbestos take a look at the images at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the images looks like a candidate for asbestos? I think only fd6, fd7, and fd8 are asbestos, and the rest of the fibrous materials imaged are cellulose binding agents used in the insulation.
I will examine the material by TEM and electron diffraction, but I was wondering if someone could post the techniques they use for sampling from the dust, and any other hints and techniques I should be aware of.
Thanks for your help, Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron MicroscopeLab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id IAA07825 for dist-Microscopy; Wed, 14 Mar 2001 08:32:38 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id IAA07822 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 14 Mar 2001 08:32:08 -0600 (CST) Received: from meta.vwh.net ([192.41.39.216]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id IAA07815 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 14 Mar 2001 08:31:56 -0600 (CST) Received: from metabolix.com (cint.metabolix.vne.verio.net [199.103.134.10]) by meta.vwh.net (8.8.5) id HAA16348; Wed, 14 Mar 2001 07:30:00 -0700 (MST) X-Authentication-Warning: meta.vwh.net: Host cint.metabolix.vne.verio.net [199.103.134.10] claimed to be metabolix.com Message-ID: {3AAF8061.D94223EE-at-metabolix.com}
I am trying to develop a procedure to thin section plant leaves, stain them with Nile blue, and visualize the stained leaves by light microscopy.
I assume that a microtome would be the best way to perform the thin sectioning but have I have no experience with microtomes. Does anyone have suggestions on suitable microtomes to purchase for such a procedure? If so, where is the best place to purchase them?
Thank you.
-Kristi Snell
-- Dr. Kristi D. Snell Metabolix, Inc. 303 Third St. Cambridge, MA 02142 Fax: 617-492-1996 Phone: 617-492-0505 x 229
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
A few years back when I took an EDS course put on by NORAN, they said keeping track of those K-alpha: L-alpha ratios should give you a running record of your detector sensitivity. I've been doing that ever since, and you can actually see those ratios change over time, as your detector window gets dirty. Another trick they mentioned (though I've never tried it myself) is to run a spectrum on some ordinary Teflon tape (yes, the kind you use for plumbing repairs). Teflon, apparently, contains no oxygen, so the presence of O in the resulting spectrum can only be from ice buildup. Clever, no? I also routinely do the Full Width Half Max test whenever I do a calibration. Good luck.
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 14, 2001 10:40 AM
I do not recall any problems with salt/water imaging.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----------------------------------------------------------------------. } } } We turned water to ice on plant surfaces using the Peltier } stage. We did not take any nice pictures (I have a few } ugly ones!). It was an idea re ice nucleation on plants } for a student project idea that we did not follow up. } } Dave } Like Dave, I've sometimes taken pictures of ice/water etc in our ESEM, but ice is such a good insulator, (I guess), the images never seem to come out well. So, I've never saved any. Also, salt/water interactions can be difficult too, since the salt really seems to want to charge a lot, to the detriment of image acquisition. Yes, you can play around with KeV and all that to minimize these effects, but it's still a very challenging "shoot" to get useable images.
Frank Thomas Geological Survey of Canada (Atlantic) Dartmouth, Nova Scotia
Brian, We work in a UKAS accredited laboratory and have a Pentafet light element detector with rotating turret device for windowless and thin window analysis. I perform a test with a Ni sample every month using the detector in its windowless mode. We monitor the ratio of the counts in the Ni L and Ni K peaks (NiL/NiL+NiK) (knowing what the value should be immediately after reconditioning). When the monthly value falls below a certain predetermined value, we simply perform the reconditioning routine. I suppose if you don't have a light element detector then the Cu standard you suggest is much better for a decent sized L line. I find that the above works very well for tracking the detector's sensitivity over a period of time. Best of luck Martin Roe
What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
Martin J. Roe Macaulay Land Use Research Institute Craigiebuckler Aberdeen Scotland UK Phone 01224 318611 e-mail m.roe-at-mluri.sari.ac.uk
I'd say some of those fibers are definitely "candidates". They are also large enough that you could analyze by PLM. If you want to do the TEM for fun or for the experience: dust samples are usually suspended in water and filtered onto a PC or MCE filter. I'm sure someone will post the appropriate reference, I can't seem to find it here.
Matt
On Tuesday, March 13, 2001 10:37 PM, Gordon Vrololjak [SMTP:gvrdolja-at-nature.Berkeley.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 } -----------------------------------------------------------------------. } } } Hello, } I was looking at some piping in our lab while sitting on the windowsill } and noticed a sign painted over. Looking closer it was a sign for } asbestos insulation that has been 80% painted over. I was told earlier } that there was no asbestos pipe insulation in the lab. I collected some } dust samples from near the fraying pipe insulation, on top of shelves, } floor, and the windowsill and imaged them in our SEM. } } Can anyone with some experience with asbestos take a look at the images } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } images looks like a candidate for asbestos? I think only fd6, fd7, and } fd8 are asbestos, and the rest of the fibrous materials imaged are } cellulose binding agents used in the insulation. } } I will examine the material by TEM and electron diffraction, but I was } wondering if someone could post the techniques they use for sampling from } the dust, and any other hints and techniques I should be aware of. } } Thanks for your help, } Gordon. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } }
This is a general site for Design News magazine; search that site for the term "lasik" to find an uncritical article featuring the inventor of the laser-vision machine now marketed by Visx. Even though it's a puff piece, it contains a great deal of info on the process.
2 recommended against the surgery, citing the risk of bad results for small gain if you have good correction now with glasses or contacts. Here are sites they cited with good info and extensive warnings of potential problems, and some additional sites I found in my own searches:
http://www.americaneye.com/bbs3/
This is a bulletin board where you can read many personal comments.
http://www.surgicaleyes.com/
This is a site founded by people who had unsuccessful laser vision correction resulting in complications. Its major point is that success defined by Snellen eye chart acuity may be inadequate.
http://wakeup.to/lasik
The site's name should tell you its slant on the topic.
http://www-psy.ucsd.edu/~mm/eyeknowwhy/index.htm
Very well-organized site, balanced viewpoint, but a bit out of date (last updated 1999). Well worth a careful reading anyway.
Detailed study results comparing PRK and LASIK outcomes. 3 years old (1998, therefore procedures done at least a year earlier). 220 eyes in the study. Interesting point (for me): 11.8% of PRK eyes and 3.2% of LASIK eyes reported a loss of 2 Snellen lines or more of best spectacle-corrected visual acuity (in English, you can't see as well as before even with glasses).
A more recent study (1999) with better outcomes. Equipment and experience makes a significant difference; be aware the technology changes fairly rapidly. 2-line loss rates from 2% to 0.6% in larger groups (1000+ and 700+ eyes).
A very good Feb. 2000 presentation of FDA pre-market approval data for several laser systems. 2-line loss is down to 0.3% with one system, but they show glare and night-vision problems reported by ~10% of patients. Remarkable Snellen acuity results: 87% 20/20 post-op without lenses.
5 respondents to my query had actually had laser surgery. All ultimately reported being satisfied with the results. 4 had no side effects at all. 3 reported good to excellent results immediately with no side effects. One (57 years old, farsighted) reported better post-op vision than previous lens-corrected vision. 1 respondent cited trouble for almost 1 year after surgery (couldn't see TEM images properly, trouble adjusting astigmatism), and said he was "initially devastated". However, he said he adjusted and/or the problems corrected themselves after a year. He can now handle all imaging tasks, and was happier overall than with glasses.
My personal decision has been to wait and see. I'm still not satisfied with the fairly high reported incidence of glare and night-vision problems, which I think are related to limited correction diameters compared to the dilated pupil of a larger-than-average eye (mine are). The changes I saw over just a few years of clinical trial results convinced me this is still evolving technology. The risks, while small, are still too high for me to accept in a purely elective procedure.
Thank you for your patience with this long-winded summary. Hope the info is useful to some of you.
Rick Mott (still reaching for his specs in the morning)
Dear Gordon, As someone in an old building with asbestos floor tiling, asbestos fume cupboard lining and old technicians who like to machine asbestos, I have looked at lots of samples of asbestos insulation. First, when asbestos is used, it is usally 30 % of the materials by volume, so it is very evident. Second it is characterized by very long, fairly straight fibres that divide down to smaller and smaller fibres, through and beyond the range of the SEM magnification. By that critierion, none of your pictures look like they contain asbestos. It helps to have looked at a sample so you can recognise it. Third, I check all suspicious fibres by EDX. The crysotile asbestos used in most insulation has a distictive Mg-Si spectrum, with a little Ca and Fe, that isn't found in most minerals. If it has that signature, then I would suspect asbestos and do the TEM. At 07:37 PM 3/13/01 -0800, you wrote: } Hello, } I was looking at some piping in our lab while sitting on the windowsill } and noticed a sign painted over. Looking closer it was a sign for } asbestos insulation that has been 80% painted over. I was told earlier } that there was no asbestos pipe insulation in the lab. I collected some } dust samples from near the fraying pipe insulation, on top of shelves, } floor, and the windowsill and imaged them in our SEM. } } Can anyone with some experience with asbestos take a look at the images } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } images looks like a candidate for asbestos? I think only fd6, fd7, and } fd8 are asbestos, and the rest of the fibrous materials imaged are } cellulose binding agents used in the insulation. } } I will examine the material by TEM and electron diffraction, but I was } wondering if someone could post the techniques they use for sampling from } the dust, and any other hints and techniques I should be aware of. } } Thanks for your help, } Gordon. 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 e-mail: mager-at-interchg.ubc.ca
Hello All, I have the opportunity to obtain a Seiko SMI8300 focused ion beam machine. I can't find out very much about it (I didn't even know Seiko made them - I suspect they are usually native to Japan). Does anyone have any useful information or experience? Could I use one to make TEM specimens?
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
One last note: of the 4 respondents who enthusiastically recommended the procedure, all were in agreement on the importance of finding a highly experienced surgeon. Complication rates are inversely related to experience.
I've never tried this either, but doubt it should work for the stated reason. A layer of ice on the detector (actually the oxygen in the ice) will act mostly as an x-ray absorber rather than a source of O x-rays. In fact, the ice will be a poor absorber for O-K x-rays because the oxygen x-ray energy is below its absorption edge.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
---------- From: Frank Thomas Sent: Wednesday, March 14, 2001 7:30 AM To: Brian Wajdyk; Microscopy-at-sparc5.microscopy.com Subject: Re: EDS detector chracterization: sensitivity/efficiency
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Brian -
A few years back when I took an EDS course put on by NORAN, they said keeping track of those K-alpha: L-alpha ratios should give you a running record of your detector sensitivity. I've been doing that ever since, and you can actually see those ratios change over time, as your detector window gets dirty. Another trick they mentioned (though I've never tried it myself) is to run a spectrum on some ordinary Teflon tape (yes, the kind you use for plumbing repairs). Teflon, apparently, contains no oxygen, so the presence of O in the resulting spectrum can only be from ice buildup. Clever, no? I also routinely do the Full Width Half Max test whenever I do a calibration. Good luck.
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 14, 2001 10:40 AM Subject: EDS detector chracterization: sensitivity/efficiency
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Fellow Microscopists, } } Fellow Microscopists, } } I want to track the performance of our EDS detector over time. We } currently } check: peak to background ratios via peak-to-tail of Mn k-alpha, } symmetry } via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha } and } L-alpha to known values, and resolution via Full Width Half Max. What I } don't have is a good way to determine sensitivity of the detector to } determine when to remove ice/hydrocarbon build up. In the past I used a } 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha } because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. } Unfortunately I no longer have this standard or the means to acquire a } new } one thanks to the slow down in the e semiconductor industry. My best } guess } would to be to clean the detector then use the k-alpha to l-alpha ratio } of } Cu. However I know that we have the best microscopists in the world at } our } disposal. Any suggestions for sensitivity tracking, or other useful } detector characterizations I may have missed, would be greatly } appreciated. } } Sincerely, } } Brian Wajdyk } Sr. Electron Microscopist } On Semiconductor } brian.wajdyk-at-onsemi.com } 602-244-4883 } }
Perhaps a Macintosh, computer guru could figure this one out.
We are capturing JPEG images from a standard (Sony, analog 470 line, NTSC) video camera using a program called Reel Eyes. The images are captured on a Mac 8500 via the built-in S-Video port and saved as Quicktime JPG's (as mandated by the Reel Eyes program). When we put these images on our server they now appear with ".bin" appended to the file name so that the new image is described as a "Application/x-macbinary". Mac folks can open the files after processing through Stuffit but PC folks are unable to deal with the files. Anyway, this is an additional step that should not be present.
We can remove this problem by re-opening each of the images in Photoshop and saving them without the Thumbnail (or preview). So, I conclude that Quicktime is always saving with the Thumbnails in place. Does anyone know of a way that we can prevent the thumbnails from occurring in Quicktime?
Any other suggestions would be most welcome.
Many thanks,
John B.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 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 ##############################################################
The problem is that your files are encoded as Macbinary (.bin) before being stuffed. Windows machines can't decode .bin files unless they have the windows version of Stuffit expander. Encourage them to get it.
To correct this you need to change a preference setting.
In Magic Menu or Dropstuff preferences, go to the "Cross-Platform" or "MacBinary" icon and choose "Never" (instead of "Smart"). There is no way to change this preference in the StuffIt Deluxe application.
The URL for this information is {http://www.aladdinsys.com/support/techsupport/mac/dlx/dlx611.html}
Happy capturing. Gordon Nord "My real job is Mineralogy"
"John J. Bozzola" 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 } -----------------------------------------------------------------------. } } Perhaps a Macintosh, computer guru could figure this one out. } } We are capturing JPEG images from a standard (Sony, analog 470 line, } NTSC) video camera using a program called Reel Eyes. The images are } captured on a Mac 8500 via the built-in S-Video port and saved as } Quicktime JPG's (as mandated by the Reel Eyes program). When we put } these images on our server they now appear with ".bin" appended to } the file name so that the new image is described as a } "Application/x-macbinary". Mac folks can open the files after } processing through Stuffit but PC folks are unable to deal with the } files. Anyway, this is an additional step that should not be present. } } We can remove this problem by re-opening each of the images in } Photoshop and saving them without the Thumbnail (or preview). So, I } conclude that Quicktime is always saving with the Thumbnails in } place. Does anyone know of a way that we can prevent the thumbnails } from occurring in Quicktime? } } Any other suggestions would be most welcome. } } Many thanks, } } John B. } } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } 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 } ##############################################################
-- Gordon Nord Small Business Network Design and Construction Macintosh and Windows - Solutions and Conflicts
Nord Consultants 20594 Cornstalk Terrace Ashburn VA 20147
Yes, I did use Hitach 4500S and 3500N to study some coated fibers in the recent past years. The coating layers were up to three.
General speaking, I think this kind of preparation needs patient and do it properly at every step as the processing goes.
The steps I did for preparing the coated fibers for SEM are 1. Cut the fibers carefully into about 1 cm length of pieces 2. Arrange them in a longitudinal direction (do not hurt them) 3. Hold one end of the fibers by using a pair of tweezers 4. Apply thin M-Bond (16) on to the fibers Since the M-bond is so thin, it will go along with the fibers and surround almost every fiber very well. 5. Leave the fibers in the air (room temperature) overnight. Do not let the wet part touch any solid surface such as metal or microscope slide. Otherwise when it cures, it is very difficult to separate them. Another way to cure it is putting the fiber with the tweezers on to a hot plate at 130 C for one hour. Do not let the wet part touch the hot plate. 6. When the fibers are cured, embed them into epoxy to make a big enough piece so that you can hold and polish it for SEM. At this step, you have to avoid trapping air in the epoxy. Especially when a fabric of fibers is going to be embedded into epoxy, you have to use a pump to get rid of air bobbles before curing. You may choose a kind of epoxy which can be cured in couple of hours in air (room temperature) or on a hot plate for about 1 to 2 hours at 130 C. The epoxy I used is a kind of mixture of two parts. 7. Then you may follow the routing procedures for polishing SEM samples, i.e., grinding first then polishing, from coarse to fine carefully.
Finally if you are going to study a fabric, then you can cut a size of 1 x 1 cm2, and hold it with a tweezers at a corner, and apply a layer of M-bond (16) on both sides, put it onto a hot plate for curing. Then embed this piece of fabric into epoxy. Again, you have to pump the air out before curing.
Good luck.
Zhenquan Liu
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Zhenquan Liu (Dr.)
------------------------------------ Zhenquan Liu (Dr.) Arizona State University Center for Solid State Science Room PSA213 Tempe, AZ 85287 Tel (480) 965 4544 (o) (480) 775 7428 (h) Fax (480) 965 9004 (o) Email zhenquan.liu-at-asu.edu ------------------------------------
Hello, We have a Gatan Duomill IBT Model 600. We have been having some problems with it lately. When I turn on the IBT, the vacuum in the main chamber works fine. When pumping down the specimen chambers, the left side does not pump at all. The right side pumps to the proper level, but when you lower the sample into the chamber, it takes a while for the vacuum to recover. There is no vacuum instability during sample rotation. There is no arcing in the guns. The argon flow is at the proper level. We have done the following: -cleaned, regreased and changed o-rings in the Whisperlock Assembly, sample viewing port, sample chamber, vent/vac buttons, and ion guns -changed the cathode tubes -dusted out the main sample chamber -check the vacuum pumps and topped off oil in the roughing pump -DP seems to be running fine
Still it doesn't seem to help. We are lost. Any suggestions about what might be going wrong here? Thanks. Prad
Pradyumna Prabhumirashi Department of Materials Science Northwestern University Phone: (847)-491-7798 Fax: (847)-491-7820 http://vpd.ms.northwestern.edu/prad.htm
We have used the k:alpha:l-alpha ratio for copper and nickel for many years as a measure of ice buildup on our detectors. Either ratio is very sensitive to ice buildup. With single element standards you don't have to worry if the mixture of two elements in your standard has changed. For sources we use copper or nickel TEM grids which are cheap and available from all the EM vendors.
Regards, Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Brian.Wajdyk-at-onsemi.com on 03/14/2001 10:24:35 AM To: Microscopy-at-sparc5.Microscopy.Com cc:
Fellow Microscopists,
Fellow Microscopists,
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
Just got a call from a researcher on campus looking for service on a Reichert Frigo-Cut type 2800, s/n 0110117 microtome. I am not familiar with this piece of equipment (we don't do light level microtomy) and I do not know this gentleman but he had already contacted one northern California service and was quoted $130/hr p to p. He balked. I explained that I had been quoted as much from services in New Hampshire. If anyone has a resource for repairing one of these microtomes in the Northern California region I will pass the info back to this person.
Thanks in advance.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Thanks for all of your replies, We had someone from our physical plant come in, and they revealed that the colour of the painted over sign indicates asbestos - free - insulation. Wierd scare for me since the painter covered the most important part of the label - that the insulation was asbestos free, not asbestos. They'll come back to tape back up all of the loose insulation tomorrow.
Since one of the specimens I looked at is very similar in morphology to asbestos, they'll be doing some wipe tests and counts by polarized light microscopy for asbestos particles to be sure. It likely was left behind from when the original asbestos insulation was removed about 10 years ago or so.
I appreciate all of the replies on the thread and supportive messages. Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Wed, 14 Mar 2001, Mary Mager 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 Gordon, } As someone in an old building with asbestos floor tiling, asbestos fume } cupboard lining and old technicians who like to machine asbestos, I have } looked at lots of samples of asbestos insulation. First, when asbestos is } used, it is usally 30 % of the materials by volume, so it is very evident. } Second it is characterized by very long, fairly straight fibres that divide } down to smaller and smaller fibres, through and beyond the range of the SEM } magnification. By that critierion, none of your pictures look like they } contain asbestos. It helps to have looked at a sample so you can recognise } it. Third, I check all suspicious fibres by EDX. The crysotile asbestos used } in most insulation has a distictive Mg-Si spectrum, with a little Ca and Fe, } that isn't found in most minerals. If it has that signature, then I would } suspect asbestos and do the TEM. At 07:37 PM 3/13/01 -0800, you wrote: } } Hello, } } I was looking at some piping in our lab while sitting on the windowsill } } and noticed a sign painted over. Looking closer it was a sign for } } asbestos insulation that has been 80% painted over. I was told earlier } } that there was no asbestos pipe insulation in the lab. I collected some } } dust samples from near the fraying pipe insulation, on top of shelves, } } floor, and the windowsill and imaged them in our SEM. } } } } Can anyone with some experience with asbestos take a look at the images } } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } } images looks like a candidate for asbestos? I think only fd6, fd7, and } } fd8 are asbestos, and the rest of the fibrous materials imaged are } } cellulose binding agents used in the insulation. } } } } I will examine the material by TEM and electron diffraction, but I was } } wondering if someone could post the techniques they use for sampling from } } the dust, and any other hints and techniques I should be aware of. } } } } Thanks for your help, } } Gordon. } 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 } e-mail: mager-at-interchg.ubc.ca } }
A few years ago, a EDS reparer explain me a test, which seems to be interresting, to check the "health" of a detector with UTW, but I never have tried it.
With a fluorine (CaF2) sample at 10 keV, You should have a ratio of one betwween F K alpha and Ca K alpha. When not, or when it diminish in the time (less F), you have ice (absorbtion effect of F by O2). (Why 10 kev? Is 10 keV not a bit high for F. It think it's a compromise between the window caracteristics and the primary energy).
Same thing with quartz (SiO2), at 5 keV, and than, if O diminish, you have oil on the window (absorbtion effect of O2 by C).
As I said, I never tried this test, the CaF2 and SiO2 samples are in my drawer, waiting for polishing. I suppose that geologist should have an advice about that, but of course they don't often work at 5 keV for EDS.
I usually use a aluminum sample holder, with a piece of carbon adhesive tape and a copper foil, and I take a spectrum with a quarter of the SEM screen on the copper, a quarter on the carbon, and the half on the aluminium,at low mag, and at 5 keV. I adust the probe current to a preset value (typically a few nA, and each time the same value) mesured on the carbon tape (less SE and BSE, better would be a Faraday cup). So I see if the sensitivity on C K, Cu L and Al K seems to diminish from on mesure to an other. In such condition you'll see that the count rate (and of course, the spectrum shape) is very sensitive to window or detector contamination (with allways the same probe current, time constant, etc.). You can imagine similar test with for exemple, Vandium L and silicon K, etc.
best regard
Jacques
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
I was asked to perform double labelling of surface antigens on leukaemia cell cultures and cord blood progenitors.
In the past I used a pre-embedding protocol for single labelling which worked quite well.
I would like to continue with a pre-embedded protocol, but both primary antibodies are raised in the same animal and I wonder how I can block unspecific labelling in that case.
(I have a protocol for post-embedded double staining with primary AB raised in the same animal, but would prefer to use that as a last resort)
So far I used a 10 nm gold F(ab)2- AB - if I use a 5nm gold F(ab)2 AB will I be able to distinguish them in the TEM?
I would appreciate your input very much.
Many thanks
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
I have a technique that I use with clients on a wide range of fibres which takes very little time.
1. Drill two or three holes through two SEM stubs placed face to face. 2. Pass the fibres through the holes. 3. Fix in place with a water based carbon solution, make sure the fibre/stub interface is well wetted. 4. When fully dry place in liquid nitrogen CARE!!. 5. When the bubbling stops lift out CARE!! 6. Place on an insulating surface and with a knife strike the interface between the two stubs - the system will fracture. 7. When dried off you have two sets of surfaces to look at in LM or SEM.
It works great on a number of differing media other than fibres, the only snag is they must not be affected by the water base.
Steve Chapman Senior Consultant Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I imagine that the problem arises when you transfer the files to the server. I'll assume that you are using some FTP program such as Fetch. In the upload preferences, make sure that you are transferring non-text data in the "raw data" format, as opposed to MacBinary, etc. Also, make sure that the FTP doesn't append a ".bin" to raw data.
Remember you still need to have a ".jpg" at the end of the filename or else the Windows boxes still won't know how to handle the file (sigh...). There are numerous utilties on the Mac to batch process the renaming of files for cross-platform compatibility.
Hope this helps, John B.
} } Perhaps a Macintosh, computer guru could figure this one out. } } We are capturing JPEG images from a standard (Sony, analog 470 line, } NTSC) video camera using a program called Reel Eyes. The images are } captured on a Mac 8500 via the built-in S-Video port and saved as } Quicktime JPG's (as mandated by the Reel Eyes program). When we put } these images on our server they now appear with ".bin" appended to } the file name so that the new image is described as a } "Application/x-macbinary". Mac folks can open the files after } processing through Stuffit but PC folks are unable to deal with the } files. Anyway, this is an additional step that should not be present. } } We can remove this problem by re-opening each of the images in } Photoshop and saving them without the Thumbnail (or preview). So, I } conclude that Quicktime is always saving with the Thumbnails in } place. Does anyone know of a way that we can prevent the thumbnails } from occurring in Quicktime? } } Any other suggestions would be most welcome. } } Many thanks, } } John B. } } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } 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 } ##############################################################
At 01:26 PM 3/14/01 -0600, John J. Bozzola wrote: } The images are captured on a Mac 8500 via the built-in S-Video port and saved as Quicktime JPG's (as mandated by the Reel Eyes program). When we put these images on our server they now appear with ".bin" appended to the file name so that the new image is described as a "Application/x-macbinary". Mac folks can open the files after processing through Stuffit but PC folks are unable to deal with the files. Anyway, this is an additional step that should not be present. } We can remove this problem by re-opening each of the images in Photoshop and saving them without the Thumbnail (or preview). So, I conclude that Quicktime is always saving with the Thumbnails in place. Does anyone know of a way that we can prevent the thumbnails from occurring in Quicktime?
The Mac's file system is slightly different than that on a PC or a Unix server. Mac files have two parts, known as forks. The icon or thumbnail image resides in the "resource fork". The regular JPEG image data that a PC expects is in the "data fork." The resource fork also holds the Mac's association between the file and the program that made it, while the PC uses the file extension for that purpose.
MacBinary files are a way of wrapping up the data and resource forks into a single file, such as for transmission via an e-mail attachment, ftp site, etc. or in your case, handy for putting on a server that doesn't speak with forked tongue.
You didn't say much about your server. If it's a WinNT server, your admin can install some AppleTalk extensions that can make PC {-} Mac file exchange a bit easier. They automatically manage the resource fork, hiding it for PC programs but allowing networked Macs to store and access it.
You also didn't say much about which PC programs you were using. I don't know of any that handle MacBinary files. (I've written a few that did, once upon a time, but they're obscure.) You may be forced to either strip the files on the Mac, or use a utility on the PC side to convert the MacBinary files to data-only files.
I don't think Quicktime thumbnails are the problem, I think it's a MacBinary problem.
Question: I have been analyzing stress around STI (shallow trench isolation) in DRAM. In general, stress is higher at bottom corner and top corner of STI, which is probed by my data. But, the strange thing is that HOLZ line split and blurring occurs in CBED pattern at the bottom of FOX (field oxide). I know that the split and blurring is the sign of high stress. And I don't think that stress is higher at the bottom of FOX. So, my question is what makes line split and blurring except stress. Have you seen that kind of phenomenon? Thank you for your answer.
If you go to our new website at www.southbaytech.com, you will find a listing of technical papers that you can request. There are several there dealing with coated fibers. Please take a look. If any look interesting, submit a request and we will send them out to you at no charge.
When you get to the site, click on Applications support then technical papers. There are a lot of papers there, so the easiest way to find the ones you are interested in is to use the "find on page" function typically found under your edit command. If you type in "fiber", you'll find some relevant papers.
I hope this helps.
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
The ion getter pump on the CM10 (12 and 20) does have a finite life and if your machine is 10 years old with the original IGP then I suspect that the pump could be finished. I assume that it will not pump down properly on the IGP but has a good penning gauge (P3) pressure.
The pump is not a standard Edwards pump as it has an extra port so that it can pump both the column and the gun. As I understand it the pump body is cut open to replace the plates and then rewelded before being leak tested and baked.
There are companies that will do this for you but you are without the pump while they do it, this may take two weeks or more. FEI should hold replacement pumps that they would swop in a day, more expensive but faster and more convenient. It depends on your budget and facilities.
Good luck, Ron
On Thu, 15 Mar 2001 14:08:46 +0100 Marco Arienti {marienti-at-tiscalinet.it} 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 no big experience with Philips microscopes, but I need some } information. } } A CM10 installed about 10 years ago have some vacuum problems. } } Looks like the Ion Getter Pump is not working anymore. } } As far as I understood the problem may be that there is no more Titanium in } the pump. } } Is possible In this one to change the only grids (like in the Leibold pumps) } or the all pump have to be exchanged? } } It is a pump manufactured from Edwards, any info about the model? } } Thanks a lot. } } } } Marco Arienti } } www.electronrescue.com } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Could you recommend a water based carbon solution?
Dave
On Thu, 15 Mar 2001 04:56:05 -0500 Steve Chapman {PROTRAIN-at-CompuServe.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 } -----------------------------------------------------------------------. } } } Hi } } I have a technique that I use with clients on a wide range of fibres which } takes very little time. } } 1. Drill two or three holes through two SEM stubs placed face to face. } 2. Pass the fibres through the holes. } 3. Fix in place with a water based carbon solution, make sure the } fibre/stub interface is well wetted. } 4. When fully dry place in liquid nitrogen CARE!!. } 5. When the bubbling stops lift out CARE!! } 6. Place on an insulating surface and with a knife strike the } interface between the two stubs - the system will fracture. } 7. When dried off you have two sets of surfaces to look at in LM or } SEM. } } It works great on a number of differing media other than fibres, the only } snag is they must not be affected by the water base. } } Steve Chapman } Senior Consultant Protrain } For professional training in SEM, TEM and EDX world wide } www.emcourses.com }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I need some info on liquid nitrogen dispensing devices. Our EM lab has four 20 liter dewars for liquid nitrogen storage. A piece of styrofoam (2 x 6) is attached to the lid. We would like to purchase a dispensing device because it would be easier to use and less waste of liquid nitrogen. Thanks in advance Karen Schlueter PWA, ARS, USDA EM Lab 1636 East Alisal St. Salinas, CA 93905 (831) 755-2847 FAX: (831) 755-2814 kschlueter-at-salinas.ars.usda.gov
As a materials scientist, I haven't had any experience doing TEM sample prep of biological samples, so I need some assistance in this.
I've been asked to look at in the TEM some dehydrated beef looking for the changes in sarcomere length etc due to the dehydration process.
I have at my disposal staining solutions of 0.5% RuO4, and 2% methanolic UA and need some procedures on how to go about preparing these specimens. (Things like cut the dehydrated sample into cubes, stain with this for how long at this temp then embed and microtome or such.)
Thanks in advance.
dz
David Ziegler U.S. Army, SBCCOM AMSSB-RSS-MS(N) Materials Science Team, SS&T Natick, MA 01760-5020 TEL: (508) 233-6484 FAX: (508) 233-5521 Email: David.Ziegler-at-Natick.Army.Mil
I am looking to purchase a sputter coater ~$5000. I currently have a Polaron E5200 that is inoperable. I do Failure Analysis on semiconductors, probably a 6 inch chamber & Au/Pd target. Can anyone in the list give me some good tools that work for them.
Anyone familiar with BOC Scancoat Six?
Thanks in advance
Jim Arnold Senior Quality Technician / Failure Analysis Honeywell - Microelectronics and Technology Center Columbia, MD 21045 email: jim.arnold6-at-honeywell.com
For organic contamination, monitoring the ratio of TiKa to TiLa is good because TiLa is absorbed heavily by organic layers on the detector window. For ice, a spectrum from say calcium fluoride is good because again fluorine Ka is heavily absorbed by oxygen containing layers such as ice.
Carl Miller ---------------------- Forwarded by Norman C Miller/RES/Raytheon/US on 03/14/2001 02:00 PM ---------------------------
"Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} on 03/14/2001 09:40:51 AM
To: Microscopy-at-sparc5.microscopy.com cc: (bcc: Norman C Miller/RES/Raytheon/US)
Fellow Microscopists,
Fellow Microscopists,
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
I am looking to replace my target on my sputter coater and would like to find out if there is an optimal thickness for the target. Is it material dependant? I had requested 2.5mil from the thickness of my old target but the vendor I spoke with will only source a minimum 4mil thick target. I am looking at Pt and Au/Pd. Will I have to increase current to get the same deposition rate for a thicker target?
TIA
David B Rose W. L. Gore and Associates 297 Blue Ball Road Elkton, MD 21921 410-506-2958
Many thanks to all of you who offered advice, assistance, consolation and even a loan of the part or a used instrument for parts. I was able to find a replacement ($100 - overpriced perhaps but worth every penny!) and will get it installed next week with the directions from another microscopy list reader.
Again my thanks to all and if anyone wants the names and addresses of some LKB parts dealers and/or service folks, I will gladly provide you with my new list off-line.
Some years ago I had Melins (510-234-5749) repair our Reichert 2800. They insisted on taking it to their shop and it needed a new compressor from Europe. The bill was over $2000. $130/ hour is a typical charge for this type of work in this area. When I the Reichert unit was destroyed in a water flood I purchased a Microm cryostat. It uses standard easily available refrigeration components. The cost and repair time is gresatly reduced. Good luck.
At 03:35 PM 3/14/01 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
No, thicker target = longer life, but it makes no difference to deposition rate, current etc. Chris
----- Original Message ----- } From: {"drose-at-wlgore.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 7:23 PM
No, thicker target = longer life, but it makes no difference to deposition rate, current etc. Chris
----- Original Message ----- } From: {"drose-at-wlgore.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 7:23 PM
G'day
I have an aging Jeol 840 and the monitor image is getting a bit dim and noisy to the extent that I have to use large spot sizes that are damaging some samples! Jeol tell me they cannot supply new tubes. Does anyone know of compatible tubes that could be utilised? Is there anything special about SEM monitor tubes?
Thanks Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
I have some difficulties to stick my protein (Mojor Sperm Protein, a basic protein from Nematode Sperm ) to the microscope slide, therefore, it is hard to perform further perfusion experiment. When I try to do so, I lose everything. Does anyone have experience in such a manipulation? Any suggestion is highly appreciated. Long
The CRTs can be rebuilt for about $600.00 to $800.00 USD by Richardson Electronics in Georgia.
I have their number & can give it to you offline.
Earl weltmer
----- Original Message ----- } From: "Dave Phelan" {emudp-at-mail.newcastle.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 4:35 PM
Try Richardson Electronics http://www.rell.com/ . Have CRT information handy (labels on the CRT) when calling Richardson Electronic. Good luck.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
G'day
I have an aging Jeol 840 and the monitor image is getting a bit dim and noisy to the extent that I have to use large spot sizes that are damaging some samples! Jeol tell me they cannot supply new tubes. Does anyone know of compatible tubes that could be utilised? Is there anything special about SEM monitor tubes?
Thanks Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
Hi: We are lookng to purchase a Zeiss 10C TEM, preferable in working condition but would consider one needing some work. Please contact us direct via e-mail or call 909 301-9130 (Los Angeles area) EMSI Peter Jordan
Dear All, I'm looking for circuit diags for J-TEC X-ray counting electronic units XM-XCU. These were supplied with some JEOL 733 'probes although most seem to have gone out with ORTEC. Can anyone help? Thanks, MAlc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
Both display and photo-monitor tubes can be recoated. We had our photo CRT recoated by Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if they're still in business, but they did a good job. JSIII
} Try Richardson Electronics http://www.rell.com/ . Have CRT information handy } (labels on the CRT) when calling Richardson Electronic. Good luck. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } } This message is made of 100% recycled electrons. } -----Original Message----- } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Thursday, March 15, 2001 10:28 PM } Subject: Jeol 840 viewing screen replacement } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Julian P.S. Smith III Dept. of Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x6427 (vox) 803-323-3448 (fax)
Does anyone use this resin and use it for antibody staining? We will be using this resin for embedding. Also any suggestions on etching. Thanks. Caroline Miller
I have a Tracor Northern TN-5402 EDS system mounted on a JEOL SEM. The monitor just went up in smoke and I need to a) find a replacement or b) try to replace the hardware. Since I don't have much money and we would like to replace the whole system in a couple of years I can't spend much to repair the monitor.
Does anyone have a monitor for sale/offer? Does anyone know of a replacement monitor available on the market? A standard BNC type of monitor does not work because of a proprietary modulation. Has anyone successfully upgraded the hardware minus the detector?
Thanks, Louie
-- Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
VISIT OUR WEB SITE: http://www.mbl.edu/ http://www.courses.mbl.edu/
Our Sorvall MT2B is having some problems, to say the least. I would very much appreciate any info. concerning service of this machine. We are located in Milwaukee, WI. I have contacted Boeckeler (new owners of RMC) regarding service. They informed me they are no longer providing service on these models. What other options do I have?
TIA Craig M. Klotz, BS,CT(ASCP) EM Tech II Neuromuscular Lab Medical College of Wisconsin cklotz-at-mcw.edu
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } 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 } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } 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 } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } 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 } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } 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 } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
Claudia, I would direct you to the March issue of journal of histochemistry and cytochemistry, they have published a procedure to double-label with gold at the pre-embedding stages, good luck Marge
Margaret Springett IEM Specialist Electron Microscopy Core Facility Mayo Foundation email: springett.margaret-at-mayo.edu
} ---------- } From: Claudia Hayward-Costa } Sent: Thursday, March 15, 2001 2:49 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: IEM: Double labelling of 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 } -----------------------------------------------------------------------. } } } } Dear Microscopists, } } I was asked to perform double labelling of surface antigens on } leukaemia cell cultures and cord blood progenitors. } } In the past I used a pre-embedding protocol for single labelling } which worked quite well. } } I would like to continue with a pre-embedded protocol, but both } primary antibodies are raised in the same animal and I wonder how } I can block unspecific labelling in that case. } } (I have a protocol for post-embedded double staining with primary } AB raised in the same animal, but would prefer to use that as a } last resort) } } So far I used a 10 nm gold F(ab)2- AB - if I use a 5nm gold F(ab)2 } AB will I be able to distinguish them in the TEM? } } I would appreciate your input very much. } } Many thanks } } Claudia } } } Dr. C. Hayward-Costa } School of Life Sciences } Kingston University } Penrhyn Road, Kingston upon Thames } Surrey KT1 2EE, UK } 44(0)208 547 2000 x 2240 } Email: c.hayward-at-kingston.ac.uk } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } 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 } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } 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 } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } 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 } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } 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 } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
As many of you know, I maintain the WWW pages for Project Micro which is dedicated to Educational Outreach to pre-college students. One of their projects is sand from around the world which is used in the classroom activities section.
Today I uploaded their latest web page documenting their sand collection administered by Joe Neilly of Abbott Labs . You can see this page at
I could not help to notice that their supply of sands from outside the USA has become sorely lacking. In particuliar
Europe, Africa and South America
are now all completely depleted. And
Australia and Asia
only have supplies from only one location left.
Can I suggest to members of this list particularly if you are not from the USA, that if you have the opportunity this would be an excellent and simple way that you can contribute to the education of our next generation of microscopist's. The sand is available to any educator, regardles of their affiliation and location.
All the information you need to contribute "sand" from your local beach is listed on that WWW page as well as the end of this message.
It will cost you a few $$ out of your pocket to send the sand , but remember, it is for a good cause. So spend a few minutes next time you drive past a beach and fill up a small plastic bag, and send it to Joe.
Cheers.... Nestor Your Friendly Neighborhood SysOp.
--------------------------- Here is an exerpt from the Sands WWW page
To Request Sand
Select the sands you would like from the list below and e-mail your request to joe.neilly-at-abbott.com. Include your selection (limit 6 per request), where the sands should be sent, and how you plan to use them. This collection was created for Microscopic Explorations but how you ultimately use them is up to you. Great stories and photos about how you used the sands are always welcome!
To Donate Sand
Sand donations are what keeps this collection going. All donations are appreciated and will be shared with many educators. To send your sand, fill a Ziplock sandwich bag half full and place it in another Ziplock bag. Mail your donation to:
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Dear Listservers, recently there was a discussion about the stability of LR-White in the EM-beam. One distribtion mentioned a polymerization of LR-White by microwave treatment. I don´t remember who sent this mail. I would be interested to test this procedure. Who knows how to do this kind of LRW polymerization? What is the set-up? How long does it take, how much Energy (Watt) is needed? Maybe the colleague who posted this mail remembers the discussion and is able to give some information.
Thanks in advance, with best regards, Michael
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________________ Alles unter einem Dach: Informationen, Fun, E-Mails. Bei WEB.DE: http://web.de Die große Welt der Kommunikation: E-Mail, Fax, SMS, WAP: http://freemail.web.de
Dear Michael: The following procedure for polymerization of LR-White has worked of me in my microwave oven: I will start with the infiltration phase: The wattage setting I used was about 700-750 watts.
1:1 95% ETOH / LR-White 50'C temperature restriction setting (trs) x2 for 10 minutes each.
100% LR-White 50'C (trs) x3 for 10 minutes each.
Polymerization in the oven: In this phase be sure to fill the beem capsule to the very top and cap off with parafilm to make a tight seal to prevent water from getting into the resin. Submerge the capsules in a water bath and place the temperature probe in the bath to monitor the temp.
The blocks will be firm and will cut very well. For more information on this technique you can contact Rick Giberson at TED PELLA CORP. USA using their toll free number, I don't know what that is in Germany! He can give you many more details about LR-White and the microwave oven. I understand that lesser wattage setting can be used to infiltrate and polymerize LR-White but I have not experimented with them as yet!
Good Luck
Ron Austin (Research Associate) Dept. of Pathology L.S.U. Medical Ct. Shreveport, LA 318-675-4775 rla-at-mindspring.com
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by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id LAA21224 for dist-Microscopy; Sun, 18 Mar 2001 11:59:42 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id LAA21217 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Sun, 18 Mar 2001 11:59:11 -0600 (CST) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id LAA21210 for {microscopy-at-msa.microscopy.com} ; Sun, 18 Mar 2001 11:59:00 -0600 (CST) X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {v03130303b6daa5368faf-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Colleagues
A number of people have reminded me about the growing problems with hoof and mouth disease around the world and that sending possible organic material from country to country should not be considered at this time.
I humbly acknowledge that you are all absolutely correct!
It simply did not occur to me to connect sand with this problem. I guess my physics background is just showing it's tunnel vision, since I think of sand as inorganic compounds and neglect to think of organic "contaminants" which might be included.
For the moment it would therfore certainly be prudent to hold off collecting and/or send and sand samples for the Micro Project, even if it can be documented that the samples have been properly sterilized. Let's wait until this potential problem is controlled. It is better to error on the safe side.
Thanks again, to everyone that pointed out my error.
Dear Microscopists, I am working with Ilex paraguariensis seeds. After dissection, with mechanical extraction of the endocarp, the micropylar endosperm is enveloped by a lignified cap. This one do not allow the cellular visualization, in clarified material (Herr´s method), of the endosperm because the yellow color of the lignin. Is there a non aggressive protocol for lignin extraction? Thank´s.
Rinaldo Pires dos Santos
Dr. Rinaldo Pires dos Santos - e-mail: rinaldop-at-uol.com.br Lab. of Plant Anatomy - Dept. of Botany - UFRGS Porto Alegre - RS - Brazil
Well, I stepped in it this time. I agreed to do some "gee-whiz" shots for my dentist of a tooth with a rather large amalgam filling. Only later did it sink in that the mercury might be a problem under vacuum and the beam. Has anybody out there done this? I'd be looking at the thing with gold coating an a fairly light beam (say 10 kV), but unfortunately, not with a cold stage. I've checked the archives and found one brave soul who said that he's done this on old amalgams. I can't really say how old this one is, but I'm pretty sure it wasn't made yesterday. Call me paranoid, but I'd like some more opinions before I do something really stupid!
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I am trying to evaluate some tools that perform automated TEM sample prep prior to FIB thinning. I am looking for some one that has experience using Sela microcleaving tools in conjuction with the TEMstation. I work with failure and material analysis in the IC industry so submicron accuracy and relaibility are important concerns of mine. Also, we will soon be facing chips with low K dielectrics which from what I have heard are extreme soft and hard to polish. I am not sure if the sawing technique they employ will be able to section this type of material.
If anyone on this list has any experience with these tools and can give me insight into their worth as a prep technique please get in touch with me.
We are interested in buying a liquid helium TEM cold stage. Has anyone had experience with it and how was it? Is there any other companies selling this apart from Gatan?
I would appreciate very much for any input on this.
Best regards Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
Does anyone have a step by step protocol for decalcifying bone for transmission electron microscopy using EDTA?
Thanks,
Kenn
Dr. Corazon D. Bucana, Ph.D. Mr. Kenneth Dunner, Jr. Department of Cancer Biology U.T. M.D. Anderson Cancer Center High Resolution Electron Microscopy Facility 7777 Knight Road, Box 173 Room SRB 1.660 Houston, Texas 77054 PH: 713-792-8106 FAX:713-792-8747
I am in need of a gas flow proportional counter (GFPC) for a JEOL 733 microprobe. Alternately, I may also need some W wire of the appropriate size to restring an existing unit. Thanks for any help/advice.
Ed Holdsworth General Mgr. SEMTEC Laboratories, Inc.
I have had good luck decalcifying bone for TEM using this EDTA protocol:
Fix 1-2 mm sized bone pieces for two days in 2.5 percent glutaraldehyde in 0.1 M Sorensen's buffer.
Decalcify on a shaker for 3-7 days in 7.5 % disodium EDTA, 2.5 % glut. in 0.1 M Sorensen's buffer. (pH the decal solution to physiological pH with NaOH.)
Change to fresh decal solution every couple of days.
Check for complete decalcification by taking before and after X-rays of the tissue.
Rinse twice in buffer
Post fix in 1 % osmium in buffer.
Rinse once with buffer, then once with ddH2O
en bloc stain for 1 hour with aqueous 3% uranyl acetate.
Dehydrate in a graded series of EtOH, infiltrate, and embed in epon.
Good luck, and I hope this helps.
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Mon, 19 Mar 2001 semcore-at-audumla.mdacc.tmc.edu-at-sparc5.microscopy.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 } -----------------------------------------------------------------------. } } } Does anyone have a step by step protocol for decalcifying bone for } transmission electron microscopy using EDTA? } } Thanks, } } Kenn } } } } } } } Dr. Corazon D. Bucana, Ph.D. } Mr. Kenneth Dunner, Jr. } Department of Cancer Biology } U.T. M.D. Anderson Cancer Center } High Resolution Electron Microscopy Facility } 7777 Knight Road, Box 173 } Room SRB 1.660 } Houston, Texas 77054 } PH: 713-792-8106 } FAX:713-792-8747 }
I'm looking for a part no longer available from the manufacturer. We have a TMC Micro-g air table, and one of the black plastic parts that holds together the leveling valve (at the ends of the table) has broken.
I can't get one of these from TMC anymore -- the whole valve kit has to be bought for $120, and the bloody things were special-made for TMC.
Does anyone perchance have any of these things that they no longer need?
Thanks to all.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hi all, I need to have TEM screens recoated and heard that Grant Scientific is the place to contact. I called 803-799-6716 (a # I found in my address archives) and got the lovely fax machine scream sound. any help? thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I have examined amalgams in the SEM in the past. There is little problem with the mercury as it is amalgamated with silver. I have even used pure mercury in the SEM as a standard for WDS with no problem. I believe mercury is used in high vacuum diffusion pumps.
Good luck!
Sam O. Mancuso Group Leader, Physical Metallurgy Special Metals Corporat