an exiting new position created for our major expansion in the micro-electronics industry.
If you have - Experience in defect review/yield improvement, - Background in lithography and CD measurement, - The ability to communicate, train, and assist users of equipment, and work in a fledging highly motivated group, - An enquiry mind with imagination and the drive to see your ideas implemented, - PhD in a related area, or equivalent experience, - Preferably experience with scanning electron microscopy, - And you would like to have your home base in the Netherlands,
then please send your C.V. without delay to the undersigned.
Renumeration and conditions of employment are attractive for the right person.
Mr. J. Jackman Philips Electron Optics Building AAE-1 P.O. Box 218 5600 MD Eindhoven The Netherlands Tel. +31 40 766548 Fax. +31 40 766164 E-mail: jjn-at-eo.ine.philips.nl
Last year we had a failure of our SUTW (super ultra thin window) on our TEM EDS detector. This is our second failure of a SUTW, although not on the same machine (a SEM) and not from the same manufacture. After talking to several people, it has become apparent that the reliability of the SUTW is not very good, although manufactures claim otherwise.
I am curious what other users experiences with SUTW windows are. In particular what are the typical down times when a failure occurs, do you take any special precautions or modify your operation procedure, would you put a SUTW (instead of a standard window or turret detector) on an SEM for instant which is vented frequently using nitrogen? What was the reason for the failure?
Does anyone have experience on P4 SPM MDT scanning tunneling microscopes and atomic force microscopes? These devices are produced by someone at an institute that used to be a part of Russian Academy of Sciences.
mail me or this listserver if you have ever heard about them, and i'll be grateful!
Can anyone tell me wich concentration PBS should be used in immune-incubations for LM and EM? We got a little bit confused because some books mention to use a 0.1M PBS solution, and some mention to use 0.01M PBS. I can imagine that using a ten-fold solution can give problems. Wich concentration is the best (immuno-fluorescence LM and immuno-EM)?
Please let me know ! Thanks
Luc Analbers
*************************************************************************** * Luc Analbers * E-mail: Analbers-at-med.ruu.nl * *************************************************************************** * Utrecht University * LLL * * Medical Faculty * LLL * * Dept. Medical Physiology & * LLL A * * Sportsmedicine * LLL AA AA * * PO-box 80043 * LLL AA AA * * Zip: 3508 TA * LLLLLAAALLLAAALLL * * Utrecht The Netherlands * LLLLLAAALLLAAALLLL * * Tel: 030 - 538911 * AAA AAA * * Fax: 030 - 539036 * AAA AAA * ***************************************************************************
Aloha! What is known about the interaction between glutaraldehyde and polysaccharides? Does glut bind to polysaccharides in a way that would cause fluorescence (beyond that which binds to the small amount of protein that is present in the sample)? From what I understand, it is known how it binds to protein and that it binds to glycogen, but that it doesn't fix soluble polysaccharides, is that right? Could anyone help us out with the chemistry beyond that? Thanks in advance!
And a Happy New Year to you all!
Tina Weatherby Carvalho Biological EM Facility University of Hawaii
I have been asked to post this by the head of my research group (John Steeds). He would be grateful if you could pass it on to anyone who might be interested.
Any replies should go direct to him at J.W.Steeds-at-bristol.ac.uk
A post doctoral research assistantship is available for research on the microstructure and processing of unique metastable Ti-Mg alloys produced by vapour mixing and condensation. The material is made at the Defence Research Agency (Farnborough, UK) and close collaboration will be required with research workers at this Research Agency.
In the as-deposited state, the material is a porous, supersaturated solid solution with a nanoscale microstructure. The research will involve hot vacuum pressing and thermal treatments to eliminate porosity and obtain an optimum dispersed phase microstructure.
The hot working will be carried out in the Mechanical Engineering Department in an Instron servo controlled press equipped with a high vacuum chamber. The microstructural studies will require the use of high spatial resolution transmission electron microscopy and associated analytical techniques in the Physics Department. Some hardness and mechanical testing will be carried out in collaboration with DRA (Farnborough) to relate the microstructure to the mechanical properties.
The person to be appointed should be able to provide evidence of proven ability at relating microstructure, processing and properties in these novel alloys whilst working in an interdisciplinary team as well as considerable experience of advanced transmission electron microscopy techniques.
For more information please contact Professor J.W. Steeds by e-mail at
Quick question! I am getting small bubbles in my formvar ciated grids. How do I get rid of these bubbles? I am not overtly introducing moisture onto my glass slides am I am using EM Sciences prepared 0.25% formvar from a new bottle!!!
I don't want to make a project out of this, so input is appreciated.
Hello Jaime and all, In the way of a suggestion, you might try a new source for your formvar solution. I had the same problem after about 20 years of coating girds with no problem. It turned out the powder (I make my own solution) I was getting was contaminated with water. I found this out when I called the supplier and asked for a bottle from a new lot and all they had was that one lot. So I ordered from a new source and the problem was resolved. It, of course, took about 2 months of hair pulling and gnashing of teeth to work through all this. I hope you can resolve your problem more easily.
Sandra Zane Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
Let me clarify my question on the interaction between glutaraldehyde and polysaccharides, and the fluorescence with confocal microscopy---
As I understand it, the people who asked me about this are actually rather happy that their specimen (whatever it is) is fluorescing so well and giving great photos. But they would like to be able to explain it. I gather they think they don't have all that much protein in their sample and so would like to explain it away by glut binding to polysaccharides. Is this plausible? Or can someone come up with another explanation?
Thanks, again, for musing on this!
It's 79 F, clear, sunny, and the surf is up. Happy New Year!
Tina Weatherby Carvalho Biological EM Facility University of Hawaii
Dear Robin, There is unlikely to be damage to the eye due either to EM use per se or to radiation--in the first place, eye tissue is not fast-growing like in- testinal epithelium, and in the second place, the radiation levels should be low. At our HVEM, for example, there is { 0.25 mr/hr (usually { { 0.25 mr/hr), and the column is monitored and interlocked to shut off the beam if the level exceeds 0.75 mr/hr at any of three positions. The most serious eye safety problem, IMHO, is exposure to the chemicals used in specimen preparation. Yours, Bill Tivol
Tina, It sounds like you are fixing plant tissue, which often is autofluorescent. An easy way to check, of course, is to look at unfixed tissue. As this is the main point of your question it seems pointless to go too deeply into gluteraldehyde chemistry. If you are interested, though, I will put you in contact with an expert ( he wouldn't forgive me for making his name too public). Gluteraldehyde reacts very rapidly with amines to form numerous products. The well known reactions are with primary amines but sulfhydral groups from cysteine and imidazole side chains of histidine also help in the cross-linking process. Theoretically, primary amino groups on amino lipids should also react with gluteraldehyde. As far as I understand it, there is little chance of gluteraldehyde cross-linking carbohydrates. That they are retained in fixed material is probably due to the cross-linking of nearby proteins which hold them in place. The best visual demonstration I saw was the addition of gluteraldehyde to homogenates of different tissues that were stirring in beakers. Liver and striated muscle gelled the instant gluteraldehyde was added. Brain gelled after a couple of hours and apple leaves were still stirring when we returned the next day! Hope this is of help to you. If it makes you feel better, in New Haven it is dark before 5 pm, there is a light dusting of snow on the ground and it is cold (below zero). What's it like in the Twin Cities anyone?
The subject of ultra thin window failures is one of extreme interest to me, since MOXTEK is the largest supplier of these windows. I am afraid that an informal survey on the Net will give warped data, since usually only those that have problems respond. We have supplied over 5000 ultra thin windows. We have a very good idea of failure rates over the first year, which is our warantee period to the EDX manufacturer. We also have some visability beyond that, because we often do failure analysis on windows for EDX manufactures as a service.
The data vary due to the fact that some microscopes are harder on windows than others. The most common failure in the field is due to particles striking the window during venting. The second most common failure is touching the window with fingers, stages, samples. etc. Very rarely does the window fail in such a manner as to be traced to defects in the manufacture. All this being said we have data showing over 7 years mean time before failure from one EDX manufacturer, and a total of 5% returns over the history of SUTW shipments from another manufacturer.
I would love to get anecdotal information from people that have had problems with x-ray windows to help us in making the window more reliable. These windows are by very nature extremely fragile. Only by the most careful manufacture, testing, and installation could the high reliability of these windows have been reached.
We are beginning a new project with bone tissue. Samples are being prepared for TEM and we have a couple of procedures for decalcification. We have not found any procedures for detecting the endpoint of the decal. process.
I would grateful for any assistance regarding decal. endpoint detection and additional decal. procedures.
Thanks for your help,
John
John S. Gardner Microscopy Lab 128 WIDB Brigham Young University Provo, UT 84602
Some time ago, Dr. Lisa Detter-Hoskin asked about edge wick experiments on a paper laminate material. She invited "any other thoughts about evaluating the morphology".
We have found that Scanning Probe/Atomic Force Microscopy yields interesting information about the surface structure of wood (and many other) fibers. Recently in our laboratory we have begun to supplement the topographic information of AFM by developing methodology for point chemical analysis. In fact, one of our early successes was in distinguishing lignin from cellulose on wood pulp fibers.
If this type of information interests you, please contact me.
DON CHERNOFF 317-251-1364 ADVANCED SURFACE MICROSCOPY FAX: 317-254-8690 6009 KNYGHTON RD. E-MAIL: echernof-at-ucs.indiana.edu INDIANAPOLIS IN 46220 Toll free: 800-374-8557 ASM is an independent analytical service and contract research laboratory.
Gernot Fuchs asked about the NanoScope III height formula, in order to scale bitmap data for display by the NS3. I have studied the NS3 data format in detail, in order to create a software package called "GP3 - Enhancement Software for NanoScope III" (by the way, this software is available for sale).
Here are some specific answers to Fuchs' questions: -The factor (Zmax * 2) is used because Zmax = 220V in the data file parameter list, but the total Z range of the piezo is 440V. -Fuchs may have neglected an important parameter from the "image list". In version 2 software, this is "\Z scale". In version 3 software, this is "\Z scale height". It is necessary to supply an appropriate value for this parameter in the file header in order for NS3 to display the data correctly. Some experimentation might be necessary to select the correct value for Fuchs' purpose--I only considered this problem going in the opposite direction.
DON CHERNOFF 317-251-1364 ADVANCED SURFACE MICROSCOPY FAX: 317-254-8690 6009 KNYGHTON RD. E-MAIL: echernof-at-ucs.indiana.edu INDIANAPOLIS IN 46220 Toll free: 800-374-8557 ASM is an independent analytical service and contract research laboratory.
We are trying to classify muscle fibers on semithin sections embedded in Agar 100 for further analyze in the electron microscope. Normally one uses toulidine blue as staining but we would like to classify the fibers of different (type IIA, IIB, I,... etc) at the light microscopic level to be able to cut out the right fiber for EM. Is there a staining method for epoxy sections that is useful for this purpose? There is some polychromatic stains but I cant recall the reference. Anyone having any ideas?
SWEDISH UNIVERSITY AGRICULTURAL SCIENCES Hans Ekwall Dept. Anatomy & Histology Box 7011, S-750 07 Uppsala, SWEDEN
We are contemplating the purchase of a liuid helium cooling stage for a TEM with a side-entry goniometer. What companies besides Oxford Instruments and Gatan manufacture LHe stages? Would those of you who use such stages please give me your opinions about your equipment? Specifically, what are the lowest temperatures reached routinely, and what is the degree of difficulty in using the equipment?
Russell Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Argonne, IL 60439 (708)252-7194
I am about to buy a dye sublimation printer and hesitate between the 3 following products. 1) Tektronix 440 (that has replaced phaser IISDX) 2) Kodak XLS8600 (that has replaced the kodak color Ease) 3) Codonics NP1600 (based on the kodak XLS8600) Our electron microscopy images are all made of grey levels and I would like to use the new monochrome ribbons that are cheaper than the coloured one. More precisely, my idea would be to work with monochrome ribbons most of the times and from time to time to use coloured ribbons. * The Tektronix seems too be well adapted to do this because it has a special ribbon tray. However, its black ribbons make green-grey pictures * The Kodak monochrome ribbon gives more grey pictures, but the exchange of the ribbon could be more precarious as there is no special ribbon tray. * Codonics will sell the monochrome ribbons in France only in April ? (Although the kodak monochrome ribbons that should be the same are already available ?)
Has anyone any comments or experience on that ? Has anyone found problem running one of the three previous printers (especially the Codonics with its special image conversion software) ?
We want to sell,l and will let go as pair to best offer a 1) TEM Lynx (Leica) Tissue Processor and a 2) Grid Leica Ultra Stainer. If interested, and your offer is serious (that is not a couple hundred bucks) I will compile offers for a week and then contact the winner and runner up. Both items were purchased new in 1990 for about $15,000 and were hardly used, mainly because the work load is not sifficien and when use we end up wasting solutions. I will include some solutions with instruments.
Please respond directly to me {Fermin-at-TMC.Tulane.edu} , NOT to the user list.
************************************************************ *Cesar D. Fermin, Ph.D \|*|/ Fax (504) 587-7389 * *Tulane Medical School /|*|\ Answ. Mach.(504) 584-2618 * *Pathology/SL79 \|*|/ Secretary (504) 584-2436 * *New Orleans, La 70 112 /|*|\ Lab (504) 5841 * *Fermin-at-TMC.Tulane.edu -} Director of Morphological Services* ************************************************************
} We are trying to classify muscle fibers on semithin sections embedded in } Agar 100 for further analyze in the electron microscope. Normally one uses } toulidine blue as staining but we would like to classify the fibers of } different (type IIA, IIB, I,... etc) at the light microscopic level to be } able to cut out the right fiber for EM. } Is there a staining method for epoxy sections that is useful for this purpose? } There is some polychromatic stains but I cant recall the reference. } Anyone having any ideas? } } } SWEDISH UNIVERSITY AGRICULTURAL SCIENCES } Hans Ekwall } Dept. Anatomy & Histology } Box 7011, S-750 07 Uppsala, SWEDEN } } E-mail Hans.Ekwall-at-ah.slu.se } Voice: +46 18 672141 Telefax: +46 18 672852 }
A relatively simple and fast procedure for polychromatic staining of epoxy sections has been published by J.Tolivia et al in Histochemistry 101: 51-55. I have not used it myself. Ask me for details if this reference is not readily available to you.
Greetings!
Herman Meekes Biological Sciences ______________ ______________ University of Missouri ---__ \ / __--- 109 Tucker Hall ------__\---/__------ Columbia, MO. 65211 \( )/ Tel: 314-882-0171 V Fax: 314-882-0123 / \ e-mail: hmeekes-at-biosci.mbp.missouri.edu /___\
I can't contribute to the glut-polysacc. discussion, but since you asked for a weather report... a very mild winter here so far, but went skating last night under the stars; temp -6 F, wind chill below -20. Cold nose, cold toes, but had the ice all to myself. Warm breezes are nice, but Winter's great too. Need more snow, though. Thanks for the review of glut reactivity.
======================================================================= Chris Frethem (612)624-4652 (voice) Cell Biology & Neuroanatomy (612)624-8118 (FAX) U of MN, Minneapolis e-mail: frethem-at-lenti.med.umn.edu
On 4 Jan 1995, Paul Webster wrote:
} Tina, } It sounds like you are fixing plant tissue, which often is autofluorescent. } An easy way to check, of course, is to look at unfixed tissue. } As this is the main point of your question it seems pointless to go too deeply } into gluteraldehyde chemistry. If you are interested, though, I will put you } in contact with an expert ( he wouldn't forgive me for making his name too } public). } Gluteraldehyde reacts very rapidly with amines to form numerous products. The } well known reactions are with primary amines but sulfhydral groups from } cysteine and imidazole side chains of histidine also help in the cross-linking } process. Theoretically, primary amino groups on amino lipids should also } react with gluteraldehyde. } As far as I understand it, there is little chance of gluteraldehyde } cross-linking carbohydrates. That they are retained in fixed material is } probably due to the cross-linking of nearby proteins which hold them in place. } The best visual demonstration I saw was the addition of gluteraldehyde to } homogenates of different tissues that were stirring in beakers. Liver and } striated muscle gelled the instant gluteraldehyde was added. Brain gelled } after a couple of hours and apple leaves were still stirring when we returned } the next day! } Hope this is of help to you. } If it makes you feel better, in New Haven it is dark before 5 pm, there is a } light dusting of snow on the ground and it is cold (below zero). } What's it like in the Twin Cities anyone? } } } } }
Hair is a difficult tissue to infiltrate and embed. We've been using an extended infiltration protocol with Embed 812. Cutting thicker than usual sections (150 nm) gives marginal results. Has anyone worked with various hair samples lately? We're looking for better infiltration and less wrinkling of the cortex and medulary areas. We'd also like to get TEM pictures of whole cross and longitudinal sections. Any ideas???
--
Darryl Krueger Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Jamie- 0.25% may be a little thin, I am used to using 0.75% formvar just my 2 cents -Mike
On Wed, 4 Jan 1995, Jaime Dant wrote:
} } } } [A } } Quick question! } I am getting small bubbles in my formvar ciated grids. How do I get rid of } these bubbles? I am not overtly introducing moisture onto my glass slides } am I am using EM Sciences prepared 0.25% formvar from a new bottle!!! } } I don't want to make a project out of this, so input is appreciated. } } Thank you for your time. } } Jaime A. Dant } jaime-at-borcim.wustl.edu } } Sorry about the typo, thats formvar coated grids. }
Just because eye tissue is not fast growing, and there is no exaggerated risk of tumors, don't neglect the possibility of cataracts. The susceptibility varies with individuals, but radiation is a definite risk factor. --John Mardinly Intel Materials Tech. Dear John, It is true that radiation can be a risk factor, and it should be mon- itored for an EM instrument. In our case, with { 0.25 mr/hr, a full shift's use of the HVEM (assuming that the beam is only on 50% of the time) will de- liver { 1 mr. Over the course of a year, this adds up to { 500 mr, the limit for non-radiation-workers. If other EM's produce as little radiation as the HVEM, the exposures will also be below the allowed limit. Of course, expos- ure to any radiation should be kept to "as low as reasonably achievable", in the words of the NCRP. I would still guess that there are many other sources of eye damage more significant than radiation exposure from an EM. Yours, Bill Tivol
Fixing shark tissue shouldn't be too difficult should it? I used 4% formaldehyde (from paraformaldehyde, in 100 mM PO4 buffer, pH 7.4) which works well for MDCK cells but swelled up the shark cells. My guess is the osmolarity is too low and needs to be closer to 1000mOm. Does anyone have any advice and/or recipes? I use this fixative to prepare cells for cryosection immunocytochemistry so that I can do light and electron microscopy on the same tissue pieces (yes 4% FA for EM doe work). I have no objection to using gluteraldehyde, acrolein or any other legal substance if it works.
Thank you to all who responded to the weather question. Brief summary: Colder than here in Minneapolis and Madison (wind chill -20F, quelle surprise). Wonderful in Sidney, Australia (how is the surf?), and only two more weeks before they get to see the sun again in Tromso, Norway. Thanks in advance, Paul Webster, Yale School of Medicine.
We have a Balzers freeze-fracture 301, not exactly the latest model but working very well, and have struck a problem with the supply of crystals for the thickness monitor, which is the model QSK 113. Balzers evidently no longer supply them -- has anyone else had this problem and found another source?
Thanks, Sally ---------------------------------------------------------------------- Sally Stowe Australian National Univ. Facility Coordinator Canberra, AUSTRALIA ANU Electron Microscopy Unit Ph 61 6 249 2743 RSBS, Box 475 Email stowe-at-rsbs-central.anu.edu.au FAX 61 6 249 4891
Message-ID: {MAILQUEUE-101.950106095638.480-at-anat.uct.ac.za} To: microscopy-at-aaem.amc.anl.gov
New Year greetings to everyone,
Weather in Cape Town Hot and windy typical for this time of year. For the surfers out there SURF IS UP.
Getting to the query regarding Hair.
We routinely look at hair samples (albino and normal). We use a standard Spurr resin which gives quite a good result. I do extend the infiltration time by not putting in accelerator and allowing the resin to infiltrate overnight at 40 degrees C. I have also obtained reasonable results with Molleneur's Epon / Araldite mixture slightly modified with the same extended time in the oven for the initial infiltration. Infiltration with the accelerator is also as long as possible at 40 degrees before embedding when using the Epon / Araldite mix.
Using this method I can get good cross and longtitudinal sections.
Hope you have some success with these.
Peter _______________________________________________________________
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- -at- -at- -at- Peter D. G. Richards -at- -at- Dept Anatomy and Cell Biology -at- -at- UCT Medical School -at- -at- Observatory -at- -at- 7925 -at- -at- RSA -at- -at- Tel: 021-406 6285. -at- -at- Internet: retep-at-anat.uct.ac.za -at- -at- -at- -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
I am working on imaging polished cross sections of portland cement particles by BE and XR imaging in the SEM. The cement powder is mixed with a resin (L.R. White) and the resin is cured. Cement particles are typically finer than 100 microns. As about half of the polished section is resin I am having difficulty with the operating conditions necessary (12kV, 5nA) burning the resin. Does anyone have suggestions for a better embedding medium? The ideal material would be stable under the beam, not allow plucking of fine particles during polishing, be fairly hard, and not be hazardous to handle or for disposal. Thank you in advance for your suggestions.
In message Fri, 6 Jan 1995 06:23:40 -0500, "SALLY STOWE" {STOWE-at-rsbs-central.anu.edu.au} writes:
} We have a Balzers freeze-fracture 301, not exactly the latest model } but working very well, and have struck a problem with the supply of } crystals for the thickness monitor, which is the model QSK 113. } Balzers evidently no longer supply them -- has anyone else had this } problem and found another source? } } Thanks, } Sally } ---------------------------------------------------------------------- } Sally Stowe Australian National Univ. } Facility Coordinator Canberra, AUSTRALIA } ANU Electron Microscopy Unit Ph 61 6 249 2743 } RSBS, Box 475 } Email stowe-at-rsbs-central.anu.edu.au FAX 61 6 249 4891 } } ------------------------------------- -------------------------------- } - } ***************
I have even an even older Balzers BA360 that I use for teaching. If you have not thrown away the used quartz crystals that had stopped functioning due to heavy deposition of platinum/carbon on them, you can remove the deposits by carefully "rubbing" them out with an unused pencil eraser (the one on the back of pencils work fine) and reuse them!
************************************************************************* M.V. Parthasarathy Professor of Plant Biology Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14850 Telephone: (607) 255-1734 Fax: (607) 255-5407 E-mail: mvp2-at-cornell.edu ***********************************************************************
You might try Streuers "EPOFIX" resin, it is avail. through most EM vendors, it cures very hard, and is recomended for non-biological materials. -Mike
On Fri, 6 Jan 1995 STUTZ-at-ENH.NIST.GOV wrote:
} I am working on imaging polished cross sections of portland cement particles } by BE and XR imaging in the SEM. The cement powder is mixed with a resin } (L.R. White) and the resin is cured. Cement particles are typically finer } than 100 microns. As about half of the polished section is resin I am } having difficulty with the operating conditions necessary (12kV, 5nA) burning } the resin. Does anyone have suggestions for a better embedding medium? The } ideal material would be stable under the beam, not allow plucking of fine } particles during polishing, be fairly hard, and not be hazardous to handle } or for disposal. Thank you in advance for your suggestions. }
I am working on imaging polished cross sections of portland cement particles by BE and XR imaging in the SEM. The cement powder is mixed with a resin (L.R. White) and the resin is cured. Cement particles are typically finer than 100 microns. As about half of the polished section is resin I am having difficulty with the operating conditions necessary (12kV, 5nA) burning the resin. Does anyone have suggestions for a better embedding medium? The ideal material would be stable under the beam, not allow plucking of fine particles during polishing, be fairly hard, and not be hazardous to handle or for disposal. Thank you in advance for your suggestions.
These vary in hardness, cure rate, clarity etc. so you can select the one that best fits your needs. If you provide your name and mailing address, I'll be please to send you specifications on these materials along with prices. I'll also send information on our other materials for cutting, grinding, polishing etc for metallography and electron microscopy.
Thank you!
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
In my previous post I mentioned that we have supplied over 5000 ultra thin windows. This should have been 'over 4000 ultra thin windows.' It just seems like like a lot more....:) With my integrity back intact, Mark Lund MOXTEK, Inc Orem UT
} I am about to buy a dye sublimation printer and hesitate between the 3 following} products. } 1) Tektronix 440 (that has replaced phaser IISDX) } 2) Kodak XLS8600 (that has replaced the kodak color Ease) } 3) Codonics NP1600 (based on the kodak XLS8600)
We did a month long evaluation of several dye-sub printers for our facility, and finally received shipment of the Seiko ColorpointPSF dye-sub last week. Our original budget was in the $7,000 range, however, once the ethernet option, memory, extral printer trays, etc were factored in, the printer camein at just over $9,000.
For our evaluation we had color confocal images as well as b&w scanned gels. We sent out the files to the various resellers for sample prints. To do an apples to apples comparison. Incidentally, Codonics was the only organization with an ablitiy to ftp image files, however, their connection appeared to be a SLIP connection requiring a tremendous amount of patience when ftp ing.
We received samples from all three manufactures as well as Seiko. Now, we do know from previous postings that the quality of the printer driver often determines the quality of the hardcopy prints. There are a tremendous number of settings when printing to the dye-sub including, linear inks, color ramps, etc, things catering more to the pubishing industry. However, as we did not have the time to get into all that, we just took whatever each manufacture printed at face value. The bulk of our printing needs are gels and images from a phosphorimager.
The interesting thing is that although the Codonics was the pricier printer with rave reviews from almost everyone, we found the Seiko noticeably better for color. For b&w, the Seiko exhibited truely dark blacks. Tecktronix & Condonics prints had either a brownish or greenish tint. The Seiko also has the added advantage of printing on cheaper thermal paper. In evaluating dye-sub to purchase try to get sample prints of your own image files. Codonics sells their printer exclusively while the other manufacturers use resellers.
Feel free to email me questions directly regarding our dye-sub pruchase. There is a reseller based in New York who was tremendously helpful in obtaining the various sample prints. Our core facility is seriously considering the purchase of another dye-sub, the Itochu Pictograph, the Cadillac (or Lexus) of dye-subs.
._____________________ | Peter J. Hahn | ------------- | Thomas Jefferson University Jefferson Cancer Institute Confocal Facility JCI-BLSB-915 233 South 10th Street Philadelphia, PA. 19107 tel : 215-955-4770 | fax : 215-923-1098 | hahnp-at-jeflin.tju.edu | -------------+-----------------+----+------+
WORKSHOP ON ELECTRON MICROSCOPY OF MATERIALS (organizers: Gareth Thomas & David Barber)
A workshop on Electron Microscopy will be held at the Hong Kong University of Science and Technology between the 19th and 25th of February 1995. The aim is to present the principles of scanning and transmission microscopy, diffraction and microanalysis, specimen preparation and applications. In addition to lectures there will be daily lab sessions on microscopy, analytical methods and specimen preparation techniques.
The HKUST e.m. facility is part of a new well-equipped research centre, with several scanning and transmission microscopes from Philips and JEOL available for demonstration and hands-on use. The workshop is being supported by many of the major manufactures including JEOL, Philips, Gatan, Oxford Instruments, and South Bay Technology. Various ancillary pieces of equipment will be for use by the participants and specialists from the companies will be on hand to discuss any problems.
In addition, once the week of lectures and hands-on practice is finished, you can relax with a free trip in a motorised Chinese Junk around beautiful Hong Kong!
As the Workshop is limited in numbers, please be prompt in applying.
For further information and registration, please contact:
Miss Alice Yuen, HKUST RandD Corporation Ltd, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Hello Netters, Since there is a discussion on epoxy for SEM, I thought I'd broaden the discussion a bit. Are there any epoxies available for making TEM cross-sections (from semiconductor materials in this case) that cure quickly and are stable during ion milling and under the electron beam? I currently use 'Devcon' - a 5 minute epoxy - but it tends to evaporate under the ion beam and isn't particularly stable in the electron beam. The best glue I've ever used is good old Araldite, which you can buy at any hardware store, but takes a long time to cure. Araldite 'rapid' seems to have the same problems as Devcon. What else is available?
Richard Beanland Dept. of Mat. Sci. and Eng. University of Liverpool, P.O. Box 147, Liverpool L69 3BX England.
One of my researchers wants to identify nerve fibers in whole mounts of Drosophila muscle preparations. Any suggestions for selectively labeling nerves? References from the 1980s use cobalt filling and/or silver nitrate, but note that muscle fiber will also take up the stain.
We would like to use both TEM and SEM. Backscatter mode for silver impregnated nerves looked interesting, but not if both nerve and insect muscle take up the label.
suggestions in this regard would be appreciated. Thanks in advance
steve
---------------------------------------------------------- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego CA 92182-0057 phone: (619) 594-4523 fax: (619) 594-5676 email to sbarlow-at-sunstroke.sdsu.edu
We are beginning a project where we will be looking at the structure of different regions of several bones that are subject to stress fractures. We are planning to bleach the samples to remove the soft tissue and then use acetone to remove any fat. We were wondering if it is necessary to Critical Point Dry the samples or just allow air drying. Are there any known changes that occur in bone material when allowed to air dry? In addition, is fixation necessary when looking at just bone? I would appreciate any comments. Thanks
Mike
--------------------------------------------------------------------------- | Michael Dunlap | lab (916) 752-0284 | | Facility For Advanced Instrumentation | fax (510) 422-2282 | | University of California | mrdunlap-at-ucdavis.edu | | Davis CA, 95616 | | ===========================================================================
There are several tests, which I found in the book: "The Preparation of Decalcified Sections" by Edward B. Brain, pub. by Charles C. Thomas, 1966. Precipitation with calcium oxalate and calcein fluorescence seem to be the best alternatives to pin pricking or X-ray. Techniques for oxalate and calcein appear in pp. 138-141. I use oxalate pptn.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On Wed, 4 Jan 1995, gardnerjs wrote:
} Hi everyone: } } We are beginning a new project with bone tissue. Samples are being } prepared for TEM and we have a couple of procedures for decalcification. } We have not found any procedures for detecting the endpoint of the decal. } process. } } I would grateful for any assistance regarding decal. endpoint detection and } additional decal. procedures. } } Thanks for your help, } } John } } John S. Gardner } Microscopy Lab } 128 WIDB } Brigham Young University } Provo, UT 84602 } } Phone: 801-378-2202 } } }
We are beginning a project where we will be looking at the structure of different regions of several bones that are subject to stress fractures. We are planning to bleach the samples to remove the soft tissue and then use acetone to remove any fat. We were wondering if it is necessary to Critical Point Dry the samples or just allow air drying. Are there any known changes that occur in bone material when allowed to air dry? In addition, is fixation necessary when looking at just bone? I would appreciate any comments. Thanks
Mike
--------------------------------------------------------------------------- | Michael Dunlap | lab (916) 752-0284 | | Facility For Advanced Instrumentation | fax (510) 422-2282 | | University of California | mrdunlap-at-ucdavis.edu | | Davis CA, 95616 | | ===========================================================================
See the many fluorescent markers for neurons made by Molecular Probes, Eugene, Oregon. They have a free very informative voluminous catalog and reference guide to all their hundreds of fluorescent probes.
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
On Mon, 9 Jan 1995, Steve Barlow wrote:
} One of my researchers wants to identify nerve fibers in whole mounts of } Drosophila muscle preparations. Any suggestions for selectively labeling } nerves? References from the 1980s use cobalt filling and/or silver } nitrate, but note that muscle fiber will also take up the stain. } } We would like to use both TEM and SEM. Backscatter mode for silver } impregnated nerves looked interesting, but not if both nerve and insect } muscle take up the label. } } suggestions in this regard would be appreciated. Thanks in advance } } steve } } ---------------------------------------------------------- } Dr. Steven Barlow } EM Facility/Biology Dept. } San Diego State University } San Diego CA 92182-0057 } phone: (619) 594-4523 } fax: (619) 594-5676 } email to sbarlow-at-sunstroke.sdsu.edu }
I know others were discussing this but I have not kept up: What do people consider to be the best but not hugely expensive film recorder for recording digital images to 4 * 5 film at about 2000 lines resolution? Details: we want to make digital composite images of up to 16 objects (bivalve shells) each digitally recorded on an ETEC SEM at about 2000 line original resolution (we do not yet have this digitizer, wanting to get the recorder specifications determined first, we are aiming toward the 4PI company, any experience with them) We also want to print color images, (probably to 35 mm film?) from beta max 3/4 inch tape (we do not yet have the beta tape reader but think we have a knowledgeable source (any recommendations welcome) We want the very highest resolution for both color and spatial detail but realize that beta is only about 700 lines original (recorded on beta tape from 3 chip high resolution cameras) We make the b/w composite SEM images by enlarging 4 * 5 negatives to the correct size, cutting out the images, pasting to black background special paper, make a tmax or plus x 4 * 5 photographic negative, also photograph text letraset independanly, combine the negative and make prints in the darkroom, no digital component. We want to get rid of all the darkreoom except the final prints from a negative (we need multiple copies) and also to be able to do the color images from a beta tape through a macintosh via photoshop (we have photoshop and a mac that probably does not have enough memory, we plan to upgrade when we know what input and output requirements are needed. Sorry abou the length but any help would be appreciated. The old XL 7000 kodak film recorder was recommended by a user but is no longer mad I like the Harris b/w dry silver printer but we need negatives to make multiple copies and it does not do color AND? it may not be high enough resolution for the composite pictures? any comments? If I get good returns of info, I will post a summary to the group. Alan Pooley Marine Science SEM lab rutgers univ 908 932 8959 x 225 pooley-at-ahab.rutgers.edu
Message-Id: {9501100024.AA15042-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: EPOXIES Orig-Author: {"John Mardinly, 5-2346, Page 322-6490, SC2-24" {JMARDINLY-at-MATTEC.intel.com} }:ddn:wpafb ----------------------------------------------------------- Another epoxy that is excellent and particularly suited for rough surfaces is Varian Torr-Seal. The stuff is made to patch high vacuum systems that will be baked up to 250C. It is strong and extremely resistant to beam damage.
Message-Id: {9501100023.AA15025-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: TEM Epoxies Orig-Author: {"Dr. R. Beanland" {beanland-at-liverpool.ac.uk} }:ddn:wpafb ----------------------------------------------------------- Hello Netters, Since there is a discussion on epoxy for SEM, I thought I'd broaden the discussion a bit. Are there any epoxies available for making TEM cross-sections (from semiconductor materials in this case) that cure quickly and are stable during ion milling and under the electron beam? I currently use 'Devcon' - a 5 minute epoxy - but it tends to evaporate under the ion beam and isn't particularly stable in the electron beam. The best glue I've ever used is good old Araldite, which you can buy at any hardware store, but takes a long time to cure. Araldite 'rapid' seems to have the same problems as Devcon. What else is available?
Richard Beanland Dept. of Mat. Sci. and Eng. University of Liverpool, P.O. Box 147, Liverpool L69 3BX Englan
I am intending to stain epoxy sections of kidney biopsies for electron microscopy and am in need of a good receipe for a PTA staining method.
------------------------- Romuald Wroblewski, Ph.D. Associate Professor Department of Pathology Karolinska Institute voice/fax:+46-8-7293597 -------------------------
I am intending to stain epoxy sections of kidney biopsies for electron microscopy and am in need of a good receipe for a PTA staining method.
------------------------- Romuald Wroblewski, Ph.D. Associate Professor Department of Pathology Karolinska Institute voice/fax:+46-8-7293597 -------------------------
To all of thise interested in where to buy Epotek 353 ND:
You can get Epotek 353 ND from:
Epoxy Technology 14 Fortune Drive Billerica, MA 01821
TEL 800-227-2201 FAX: 508-663-9782
You can also buy it from us in smaller quantities - albeit a higher price!
P/N 0714-010 price: $50/8 oz kit
The only advantage in buying from us is that you can combine it with your other supply orders and we also can ship it to you the same day. Otherwise, you can get a lot more for your money buying it direct from Epoxy Technology.
I hope this helps!
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
We are finalizing plans for a new central microscopy laboratory. In part, these plans are modeled after those furnished by R A Harris (University of California, Davis) where areas are compartmentalized i.e. cryo microtoming in separate room, etc). For general health reasons we have restricted solvent usage to one room within and isolated from this larger room. We are experiencing some resistance to this concept. Can any of you lend some support to this concept? Or tell us why it is not a good idea. Please contact me via email, fax or phone with your welcome comments. As ever, thanks in advance for your comments.
L E Porter Phone (216) 796-1620 Head of Microscopy Fax (216) 796-3304 The Goodyear Tire & Rubber Company EMail LPORTER-at-GOODYEAR.COM Dept 415A 142 Goodyear Blvd Akron, OH 44305 USA
Epoxy Technology 14 Fortune Drive Billerica, MA 01821
TEL 800-227-2201 FAX: 508-663-9782
You can also buy it from us in smaller quantities - albeit a higher price!
P/N 0714-010 price: $50/8 oz kit
The only advantage in buying from us is that you can combine it with your othe r supply orders and we also can ship it to you the same day. Otherwise, you can get a lot more for your money buying it direct from Epoxy Technology.
I hope this helps!
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
I think I'm jumping into the middle of a conversation, but here are my two cents. I've used Torr-Seal for making cross sections in the past and had reasonably good luck. It takes about 24 hrs. to cure and stands up well in the ion mill; I, however, am not sure about the particle issue mentioned. My sample was well characterized before XTEM.
I still like LR White Hard Grade resin for microtoming particulate material and free standing optical thin films for XTEM. My samples are typically ceramics and metals. LR White is stable in the electron beam assuming it is C-coated and can be cured very quickly with an accelerator or in an oven. Must be mindful of potential brittleness.
Jim Heuer GE (510) 862-4501 heuerj-at-vncpo1.ne.ge.com
I think I'm jumping into the middle of a conversation, but here are my two cents. I've used Torr-Seal for making cross sections in the past and had reasonably good luck. It takes about 24 hrs. to cure and stands up well in the ion mill; I, however, am not sure about the particle issue mentioned. My sample was well characterized before XTEM.
I still like LR White Hard Grade resin for microtoming particulate material and free standing optical thin films for XTEM. My samples are typically ceramics and metals. LR White is stable in the electron beam assuming it is C-coated and can be cured very quickly with an accelerator or in an oven. Must be mindful of potential brittleness.
Jim Heuer GE (510) 862-4501 heuerj-at-vncpo1.ne.ge.com
We are interested in cross-sectioning devices on silicon with a thin layer of polyimide. This is primarily for optical or SEM imaging, not TEM. The polyimide layer is on the order of 10 micrometers thick (and possibly an order of magnitude higher). My limited microtomy experience has been that cleaving the polymer introduces substantial dimensional distortion. Does anyone have a good idea for this? We are considering trying a "standard" mechanical polish but have serious reservations because of the polymer. We also know it can be done, having seen good pictures (lacking, of course, specimen prep information).
Any suggestions will be welcome.
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu
} } Happy New Year } } I am intending to stain epoxy sections of kidney biopsies for } electron microscopy and am in need of a good receipe for a PTA staining } method. } } ------------------------- } Romuald Wroblewski, Ph.D. } Associate Professor } Department of Pathology } Karolinska Institute } voice/fax:+46-8-7293597 } ------------------------- } } Hi Romuald, I think you should try a modification of Stempak & Ward 's (1964) receipt which involves a methanolic solution of Uranyl Acetate. Just replace the UA by PTA and carefully follow their method, which is described in J. Cell Biol. 22 pp. 697-701. I have used this with success for staining proteinaceous structures embedded in Araldite. It cannot be used for material fixed in Permanganate, but this should not affect many people nowadays! Hope you have success with it. Jim Chalcroft
I just realized that you have a materials microscopy lab. Sorry - I know some of my ideas and experience is not applicable. Hope some of it is.
Good luck with the new lab.
Dan
On Tue, 10 Jan 1995, LE Porter wrote:
} We are finalizing plans for a new central microscopy laboratory. In part, } these plans are modeled after those furnished by R A Harris (University of } California, Davis) where areas are compartmentalized i.e. cryo microtoming } in separate room, etc). For general health reasons we have restricted } solvent usage to one room within and isolated from this larger room. We } are experiencing some resistance to this concept. Can any of you lend some } support to this concept? Or tell us why it is not a good idea. Please } contact me via email, fax or phone with your welcome comments. As ever, } thanks in advance for your comments. } } L E Porter Phone (216) 796-1620 } Head of Microscopy Fax (216) 796-3304 } The Goodyear Tire & Rubber Company EMail LPORTER-at-GOODYEAR.COM } Dept 415A } 142 Goodyear Blvd } Akron, OH 44305 } USA } } } }
On Tue, 10 Jan 1995, ROBERT K. NAUMAN wrote: } Can anyone provide me with a source for a new 450 Watt Xenon Lamp? } Thanks! } Bob Nauman } Baltimore, MD 21201
May contact Ralph Maxwell in Massachusetts -at- (508) 692-4973. He operates a company called Remtron that we used to retrofit our old B&L Research I Metallograph from carbon arc to Xenon. He sells the OSRAM XBO 450W OFR for about $480 per bulb. If this is not the correct type you need, I'm sure that he can help ya find the exact type. Also, if memory serves me, (haa haa!) I think OSRAM may have a facility in New York or somewhere in the east; altho' the bulbs I have are German-made.
Hope this is of help, -Rob ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /\v/\ | | Metallographic Lab. | Missouri Speleological Survey /\v/\ | | Rolla Research Center | Bat Conservation International /\v/\ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-rorc.usbm.gov | National Speleological Society #32993 /\v/\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
necessary to know what kind of examination is to be done. We have examined bone and tooth here at our facility at the U. of P. dental research department and for quick examination of both these mineralized tissues air-drying has been sufficient. However, we also first examine samples at 10x to 100x in a dissecting microscope while fresh from the source for micro-fractures to determine if any small cracks we may see in the SEM have been caused by the drying method or were there at the beginning. To minimize crystal metamorphosis, the mineral sample is immersed in LN2 and lyophilized. For a moderately rigorous drying procedure, we do a slow dry out of Freon 113 and omit critical point drying. This is done by treating the mineral as any soft tissue--Karnovsky's fixative, followed by an ethanol drying series and, after two exchanges in 100% ethanol, samples are immersed in Freon 113. The container with the bone and freon (usually the bottom of a 30 mm petri dish) is covered with parafilm with a few needle holes in it and allowed to air-dry (a few hours). An excellent reference is "Methods of Calcified Tissue Preparation" Elsevier Science Publishing Co., Inc. 52 Vanderbilt Ave., New York, NY 10017. Particularly, Chapter 7 by Alan Boyde.
I have no knowledge of the design plans you mention, but I assume that the solvent usage room has special ventilation and/or hood space. I would also assume that all the tissue embedding, grid preparation, slide staining, solution preparation, and tissue fixation (implying the presence of transmission and dissecting light microscopes) would be done in this room. If so, the only objection I can think of would involve the occasional use of small amounts of solvent (ethanol, acetone and xylene) needed for equipment cleaning, slide coverslipping, and possibly section mounting in the microtomy area. The necessary amounts of these things would be very small and it is doubtful that their presence and use in the microtomy room would constitute a hazard. If a large number of slides need to be coverslipping at one time, it is, of course, necessary to do it in a highly ventilated environment - but coverslipping an occasional individual slide shouldn't require the inconvenience of moving to another room.
If you plan to separate working with tissues (fixed or otherwise) to another room, you may generate many inconveniences unless small amounts of certain chemicals (primarily fixatives, eg. aldehydes) are allowed.
I have worked as a microscopy technologist for approximately 17 years in a number of settings. I am aware of the safety issues but, also know that if you place unrealistic restraints on people, they will find a way around them sooner ar later. The best solutions seem to me to usually involve a certain amount of compromise.
Dan
On Tue, 10 Jan 1995, LE Porter wrote:
} We are finalizing plans for a new central microscopy laboratory. In part, } these plans are modeled after those furnished by R A Harris (University of } California, Davis) where areas are compartmentalized i.e. cryo microtoming } in separate room, etc). For general health reasons we have restricted } solvent usage to one room within and isolated from this larger room. We } are experiencing some resistance to this concept. Can any of you lend some } support to this concept? Or tell us why it is not a good idea. Please } contact me via email, fax or phone with your welcome comments. As ever, } thanks in advance for your comments. } } L E Porter Phone (216) 796-1620 } Head of Microscopy Fax (216) 796-3304 } The Goodyear Tire & Rubber Company EMail LPORTER-at-GOODYEAR.COM } Dept 415A } 142 Goodyear Blvd } Akron, OH 44305 } USA } } } }
If you are using a Nikon power supply, Nikon recommends that you DO NOT USE Osram bulbs. For some reason it drastically shortens the life of the power supply (i.e. shorter than the life of the bulb!). In our case our Nikon supplier suggested Ushio bulbs.
We specifically have a 75 watts system and the same may not be true of the 450 watt systems but you may want to find this information out before you fry your power supply.
(I have nothing against Osram, I regularly use Osram bulbs for many other applications)
Richard E. Edelmann Electron Microscopy Facility Supervisor Marshall University - School of Medicine Huntington, West Virginia
keeping the solvents "restricted" to the prep lab is a good idea, however in reality it is sometimes necessary to us solvents (chloroform, acetone, ethanol) in thet microtomy or microscope labs. -Mike
Is anyone knowledgeable about Tracor Northern NS-880 EDS x-ray analysis = system? Specific question: How do you boot from the tape drive when the = system has had a floppy drive but it is missing?
Any information would be greatly appreciated. Dr. N.K. Smith
} Does anyone know where to buy freon 113 in gal. size? Our freon pump } cooling the Cameca MBX probe needs periodic refill. } } Thanks in advance! } } Vincent } } PS. I really doubt that the heavy molecules like freon 113 can fly } high to the ozone layer. Most likely they "sink" to the ground level } and help fighting the ozone created by industrials and autos(?).
Heuer Jim P. writes:
} Try "Fluorinert" Electronic Liquid (FC-77) from 3M Co. (213) 726 6361. } About $300 for 11 lbs. (3/4 gal.)
I'd suggest a different 3M material, SF-2, which is used as a blanket material for vapor phase soldering. It might be a better replacement, but that all depends on what you mean by a Freon pump. Is the material used as a heat transfer medium or is it evaporated? If the material is used in a closed system, the FC-77 would be an adequate choice.
Tom Ostertag ostertag_tom-at-mn15-gw.mavd.honeywell.com
We have a user with a nasty problem that we haven't been able to dent. They are trying to do some immunocytochemistry on HeLa cells growing on glass coverslips. They fix in either acetone or 2% paraformaldehyde. The paraformaldehyde fixed monolayers are subsequently permeabilized with acetone, triton x-100 or saponin (problem is the same regardless). At this point there is no autofluorescence at any wavelength. When they incubate the cells in a high quality donkey anti-mouse IgG coupled to Cy5 (Jackson) there is a very high level on non-specific adhesion. We have tried the following blocking agents: normal Donkey serum, BSA, gelatin, non-fat dry milk, or all of the above simultaneously. Pretreating the sections with the blocking agents has no effect. Using other anti-mouse antibodies coupled to FITC or rhodamine or a Goat anti-rabbit serum give the same problem. I should point out we routinely use these antibodies in my own laboratory following the same protocols and do not get any background binding. We get the same result when we personally run up the cells so we can't blame it on the technician. Is there something weird about HeLa cells or something else I am missing? Do HeLa cells have IgG receptors? Your helpful advice will be gratefully received.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
Fellow Researchers, If anyone knows of research being done on bowel disorders I have a sick daughter and her doctors are at the brain-storming stage after a year of exploring. This is a personal request, please contact me through my E-Mail address. I appreciate any help at this point. Thnaks.
Someone here in our lab also uses HeLa cells, and stains tehm for coxsackie virus using monocloanl antibodies. He has not had any problems with sticky secondaries. He did say that HeLa cells can possess/express Fc receptors. These might be isotype specific depending on the "strain" of HeLa cells you have. Otherwise can't think of any other reason, but what concentration of blocking agents did you try?
Mark Elliott Pulmonary Research Lab UBC Vancouver Canada
Are you using Fab fragments for your secondary antibodies? Generally, if we have problems it is because we are using the whole antibody. Also going to affinity purified or the highest specificity we can obtain usually gives the best results. The addition of serum (ie goat serum for goat anti-mouse IgG) throughout the incubation is also helpful.
_____________________________________________________________________ | | | | James V. Jester | Dept Ophthalmology | | jester-at-crnjjsgi.swmed.edu | UT Southwestern Medical Center | | TEL (214)648-7215 | 5323 Harry Hines Blvd | | FAX (214)648-2382 | Dallas, TX 75235-9057 | |__________________________________|__________________________________|
Does anyone know the name and contact information for a person at UC = Irvine who had three Tracor Northern NS-880 x-ray analysis systems? This = is as per John Liska, formerly with Tracor Northern--Noran. Information = is greatly appreciated,,,Dr. N.K.R. Smith
In regards to the response concerning xenon bulbs, I'd like toprovide the following information.
Nikon's 75 watt xenon power supply is usually shipped with Osram 75 watt arc lamps and that is the recommended supplier, although technically there is no difference between Osram and Ushio.
Nikon's 100 watt xenon power supply sold outside the USA is shipped with Ushio 100 watt xenon arc lamps. I believe only ushio manufactures this type of 100 watt arc lamp.
In Nikon's 100 watt mercury power supply either Osram or Ushio 100 watt mercury arc lamps of the same specification are usable and performance is the same.
450 watt xenon bulbs were typically used in Microprojectors used to project microscope slide images on to a projection screen for use in teaching, CCTV has now replaced this technique.
Regards,
Stan Schwartz Manager, Biomedical Instruments Nikon Inc. Instrument Group 1300 Walt Whitman Rd. Melville, NY 11747 516-547-8529 Fax 516-547-0306 E-mail NIKONBIO-at-aol.com
We use CD ROM for archival image/data storage. CD Writer's have come down in price quite abit lately. What I originally paid $5K for is now available for ~$2K and the unit is twice as fast. You may want to seriously look into these instead of tape. Blank CD's cost ~$15 and hold ~650Mbytes.
There are several vendors on the market. I am currently using Pinnacle Micro, but there are at least 2 other manufacturers out there.
A postdoctoral position is available (immediately) exploiting the unique capabilities of a UHV-HREM (2 Angstroms and 6x10-11 torr) at Northwestern. Connected to this instrument are a number of analytical (XPS, Auger, SEM) and a GaAs deposition chamber. The position would involve working on projects involving GaAs deposition, as well as number of other projects involving small particles and metal-semiconductor surfaces.
Experience with TEM (HREM preferred) plus surface science techniques are important.
Send C.V. plus other material to:
L. D. Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208 ldm-at-apollo.numis.nwu.edu
We want to set up a computer-based index to our EM lab negative files. This is a request for your experiences and recommendations of software. We would like to store and be able to search thumbnail views of each image as well as assignable fields and keywords. The images themselves will be kept as negatives, positives, slides and other formats as separate files. Commercial packages which manage images themselves are reported to suffer in searching speed, power and reliablity (A.Abernathy, "Managing your media", MacUser Sept. 1993, Pp. 190-206). Perhaps a database program that allows thumbnails to be imported would be best. Your comments invited.
jacob Jacob Bastacky, MD 1-116 Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
I for one would greatly appreciate the names of the other two companies (with telephone numbers if possible). Also, what is your opinion and experience with the Pinnacle Micro? Any recommendatins?
On Fri, 13 Jan 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} Keith } } We use CD ROM for archival image/data storage. } CD Writer's have come down in price quite abit lately. } What I originally paid $5K for is now available for } ~$2K and the unit is twice as fast. You may want } to seriously look into these instead of tape. } Blank CD's cost ~$15 and hold ~650Mbytes. } } There are several vendors on the market. I am } currently using Pinnacle Micro, but there are } at least 2 other manufacturers out there. } } Nestor Zaluzec } ANL }
Multiple Write sessions on a single CD-ROM have been possible for several years. The problem is that many inexpensive CD ROM readers donot support Multi-session formated disks. So if you intend to send/give the data to someone with a standard format CD reader (i.e. the run-of-the-mill ~$200-300 variety) you will have to cut them a new disk. This generally is not a problem assuming that you already have the CD writer.
Since the whole reason for a visitor to come to a user facility is to do an experiment and leave with the data I see no conflict with not "filling" up the disk. For example if you are in a user facility and a visitor comes to do some experiments collecting a few hundred Mbytes of data, then writing a single CD with just that few 100Mb on it is usually more effective than writing tens of floppies or several Syquests. The logic to a CD vs a removable/rewritable media (Syquest,MO, etc..) is that everyone can generally afford to buy a $200 CD reader but not everyone can afford to buy a ~$900 MO reader. This way a "user-facility" can support many type of users and they do not all have to go out and buy expensive drives. On the flip side if the user and the facility both have compatible media then using that media is reasonable (since I happen to have lots of different drives and media I can accomodate most users)
For local storage issues, the approach I tend to take is to have a local disk server (2.5 Gb partitioned into 3-650 MB segements plus a scratch disk) which is used to temporairly store data . When a disk partition is full then it is written to CD (usually 2 copies) and then cleared. If an individual user wants to archive things this procedure can be also used. In this case, we download his/her data overnight to an empty partition and then write the data and give the user back the finished CD. Then wipe the partition for the next user.
Generally I tend to avoid multi-session CD's, but have used them on occassion. You just have to carefully label the disk. Also when you mount a multi-session CD you get alot of "virtual" disk icons mounted on your desktop. One per CD session. So if you get into the habit of writing small partitions to your CD then you will find lots of icons cluttering up your desktop. My real desk is bad enough so I try to avoid the problem propagating to the screen of my workstation.
Hope this proves useful...
Nestor
--------------------- P.S. Someone asked for information on phone numbers for manufacturers of CD writers I pulled this out of MacWorld Magazine.
Pinnacle Micro RCD-1000 CD writer: 800-553-7070
Most CD writer's are sold direct by the manufacturer I don't recall seeing very many advertised in the mail-order catalogs. So you may have to do some hunting. I'm positive that there are at least 2 other manufacturers out there of ~$2K writers. I seem to recall Sony and Chinon, but don't take that as Gospel.
BTW for the record I have no financial interest in Pinnacle Micro Systems.
I use Claris Works database to maintain my records and include film numbers for each record or subject. Any search for an image is slowed by division into years but I have done retreivals rapidly by using one criterion for search. Good Luck-- Kate Connolly
Like Nestor: We have stopped using tape completely,(anyone want a Jetstore 5000!) having found that we actually needed our backup on occasion. We are using the Pinnacle RCD1000 currently, it cost -at-$2000 to go in a pc, this included an Adaptec SCSI board. Generally we generate about 1 Gig/mo of images from a variety of platforms, Confocal, EM, and LM. However I would like to make a few comments on the viability of CD's as a principle storage media
1: They are an absolute boon in a multiuser environment. We give users a CD and they are gone forever, we do not have to coddle their data for the next 20 years or try and find it on a tape It doesn't matter what platform they use,(PC/MAC/SGI) the CD works. However this is tempered by two problems. a: The recordable CD's are not as robust as ones made by commercial presses and do not like being scratched. b: The namespace of the platform chosen is important. For example we use SGI's to analyse all our confocal data however the data may be stored on a Novell server and accessed through NFS. When the data is archived to CD, only the DOS name space is recorded, which means that some of the UNIX names get truncated to 8 letters and then become useless
2:Cd's are geat if you have 650meg to back up. One of the biggest problems we faced when we started using CD's was how to organize the data. By project? by date etc. The problem is that unlike a tape or disk where you can keep adding on forever with a CD you loose a lot of space when the disk is multisessioned. Effectively the FAT takes up 20MEG and every time you add something to the disk in a new recording session you have to rewrite the FAT.... To get round this problem as we ideally we would like to make single user archives we have a large online resevoir of space (13MEG) and use 650MEG Flop-opticals as an intermediate storage device.
3:You need to buy a 1 Gig drive along with the CD. It is possible to make a CD with a virtual image which then pulls all the data from the vaious directories and drives etc. However this is dangerous... A much better solution is to plant a decent sized hard drive in the machine you will be archiving and make a single, large ISO compliant file which is written directly to the disk.
4:When recording you cannot expect the pc to do anything else. This includes running a screen saver. I estimate that the happy Xmas Screen saver we had this year cost us about 5 CD's.
5:It is worth thinking about a Juke box player to go with it. We are using a Pioneed 604X Takes 6 disk cassettes, costs about $1000. Trouble is that if you want to use it over a net then you must buy multidisk support software/hardware. Microtest make either diskport (a hardware soln) or diskserve (a software soln). Either work well.
Generally we are extremely happy with this media, We have Flop-opticals, DAT and non-DAT backup devices. CD's are really the only way to go though they are cheap ($15/650meg) relatively fast and perhaps most importantly now that almost all PC's have CD players in them, distributable!
---------------------------------------------------- Simon Watkins Ph.D Director Structural Biology Imaging Center Scaife 840 University of Pittsburgh Pittsburgh PA 15261
} We want to set up a computer-based index to our EM lab negative files. } This is a request for your experiences and recommendations of software. } We would like to store and be able to search thumbnail views of each } image as well as assignable fields and keywords. The images themselves } will be kept as negatives, positives, slides and other formats as separate } files. Commercial packages which manage images themselves are reported to } suffer in searching speed, power and reliablity (A.Abernathy, "Managing } your media", MacUser Sept. 1993, Pp. 190-206). Perhaps a database program } that allows thumbnails to be imported would be best. } Your comments invited.
The new version of HiJack Pro solves many of these problems in a cost effective ($90) fashion. It works differently to the earlier version and keeps a central database of images which can be accessed very rapidly. It works accross servers, allows multiple indexes, does thumbnails, and updates itself automatically, comes with a montaging and editing program. We love it
---------------------------------------------------- Simon Watkins Ph.D Director Structural Biology Imaging Center Scaife 840 University of Pittsburgh Pittsburgh PA 15261
Hi, I am currently looking into switching to digital image archiving and have found two referances which may be of assistance to others in the same position:
- There is a review of cd-rom drives in "PC Computing", Nov., 1994. This supplies name and phone number for several manufacturers as well. - Kodak offers a CD-ROM drive guide which lists CD-ROM drives, controller cards and device driver software combinations which are compatabile with Kodak Photo CD. Not all combinations are equal. I have publication DCI-249 dated 8/94 which lists combinations for PC and Mac. I don't know what is available for Unix.
Hope this is helpful. Laurie
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
14th International Congress on X-ray Optics and Microanalysis August 29 - Spetember 2nd 1995 GuangZhou, CHina
This congress will be held, 39 years after the first UK congress, in GuangZhou, a city only two hours by train from Hong Kong. The program will include the following topics: X-ray Optics, Sources and Microscopy; X-ray Photoelectron Spectroscopy; Analytical Electron Microscopy (AEM); Scanning Electron Microscopy; Electron Probe Microanalysis; Auger Electron Microscopy; Ion Probe and Secondary Ion Mass Spectroscopy; Laser Probe; Scanning Probe Microscopies (STM, AFM); and Confocal Microscopy.
Contributed papers prepared in 2 or 4 camera ready pages with text and figures inside a frame of 144mmx210mm will be published in Proceedings available to each participant at the Congress. A comprehensive Trade Exhibition will also be scheduled.
The deadline for contributed papers is April 30th, 1995, and the early registration fee, thru June 30th, is US$350. The accomodation rate (including three meals) in a three star hotel is US $50-85 per day. The organizing committee will provide arriving participants with transportation from GuangZhou International Airport and the railroad station on Aug.29th.
Second circulars, registration forms and contributed paper format are now available from
On the subject of CD ROMS writers, I looked through a British computer magazine and found advertisments for a few manufacturers, of the ones likely to be available in the US try Philips (UK price ~$3000) or Yamaha who manufacture a quad-speed writer (~$5000). However prices in the UK tend to be much higher than in the US, so they could be considerably less over there. I hope this is of use to somebody!
Doug Arrell +------------------------------------+ | Dr Douglas Arrell | | Mechanical Performance and Joining | | Institute for Advanced Materials | | 1755 ZG Petten | | Netherlands | | {ARRELL-at-JRC.NL} | | Tel. (+31) 2246 5287 | | Fax (+31) 2246 1917 | +------------------------------------+
I am in need of optical crystallographic properties for FeCl3 and FeCl3 - nH20. Winchell's Optical Properties of Artificial Minerals has the +2 valence state of iron, but not the +3 state.
The June 94' issue of MacUser has a good article on CD-ROM writers listing a number of different companys and MacUser's ideas on the pros and cons of each. Included is MacUser's commemnts on different software that is available for the writers. I beileve PC Week had a simular article.
Mike
--------------------------------------------------------------------------- | Michael Dunlap | lab (916) 752-0284 | | Facility For Advanced Instrumentation | fax (510) 422-2282 | | University of California | mrdunlap-at-ucdavis.edu | | Davis CA, 95616 | | ===========================================================================
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu Time: 4:39 PM
Date:1/16/95 URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html NC EMAL
Three software packages have been added/modified in the Microbeam Analysis Society Software Library (MASSL) in recent months. The MASSL resides on freebie.engin.umich.edu username: anonymous password: your email address in the form user-at-domain. Directory: /pub/MSA+MAS/MASSL New files in: 1. /pub/MSA+MAS/MASSL/xphi A Full abstract was not included by the author of this program. Here is a summ ary of the information that he supplied in printed form.
Xphi is a PhiRhoZ correction program. For a description see: "An Accurate Computer Correction Program for Quantitative Electron Probe Microa nalysis", Claude Merlet, Mikrochim. Acta 114/115 363-376 (1994). Install the program by exploding the auto-unzip file xphiz.exe into a directory on your hard drive (say, c:\xphi). Install the fonts, FIX8X14.FON and FIX8X16.F ON that come with the program in the Windows sytem directory (e.g. in c:\windows \system). The program is called xphi.exe, add the program to your program manag er listing so that you may double click it to run it from the windows shell.
2. /pub/MSA+MAS/MASSL/shrli A program called Simply Shrli, designed to run on the PC. This from the readme file. SIMPLY : WHAT IS IT FOR ? ************************* SIMPLY is a set of programs dedicated to Transmission Electron Microscopy in
Materials Sciences. It allows you to BUILD structures, DRAW and HANDLE cells, CALCULATE, DISPLAY and HELP to INDEX diffraction patterns, CALCULATE and DIS-
PLAY High Resolution IMAGES. SIMPLY runs "SHRLI", (c) M.A.O'KEEFE, (1980). .../...
SIMPLY first appeared as: "SIMPLY: a package for the SIMulation and disPLaY of HREM images on PCs", T. EPICIER, M.A.O'KEEFE, communication at 33rd Ann. Meeting of the French Electron Microsc. Soc. (SFME), Villeurbanne-Lyon - F., june-july 1993. It has been demonstrated during the "Computers Open lab", at the 13th Intern. Congress on Electron Microscopy (ICEM 13), Paris - F., 17-22 July 1994.
3. /pub/MSA+MAS/MASSL/KAKER Two abstracts here in the requested format (MANY THANKS Henrik!!!). Title : Edax Keywords : XEDS,SEM Computer : IBM PC or compatible Operating System : MS-DOS Programming Language : Turbo Basic,GWBasic Hardware Requirements : EDS Edax 9100,RS232C Card Author(s) : Henrik Kaker Correspondence Address : SEM/EDS Laboratory,.Metal d.o.o.,Koroska c.14 : 62390 Ravne,Slovenia,E-mail:kaker&ctklj.ctk.si Abstract:
Program for connection energy dispersive spectromter Edax 9100 with IBM PC or compatible computer via line printer port of the Edax 9100.Program allows transfering intensity and spectral data to PC and processing this data with standardless programs for bulk and thin film samples.
Documentation: EDAX.DOC
Source Code: Yes
and
Title : EPMA Database Keywords : EDS, WDS, SEM, EPMA Computer : None Operating System : None Programming Language : None Hardware Requirements : None Author(s) : Henrik Kaker Correspondence Address : SEM/EDS Laboratory,.Metal d.o.o.,Koroska c.14 : 62390 Ravne,Slovenia,E-mail:kaker&ctklj.ctk.si Abstract:
Database of 681, 826 and 40 published entries for performance testing of EPMA programs. Database EPMA40.DAT present collection of published data from different manufactures of EDS and WDS hardware and software and is suitable for testing standardless (no-standard) analysis programs.
Documentation: EPMA.DOC
Source Code: No
Two other points of information. A. Some of the archived files have been found to be corrupted and even our backups are no good. If anyone has good copies of the following files, please send me copies. They are under pub/MSA+MAS/MASSL 1) ~/ECPORI/ECPORI.SIT 2) ~/FINDCSL/FINDCSL.SIT B. The MASSL and the Electron Microscopy and Microanalysis Public Domain Libraries will be merged into one library, however this will only occur when Nestor Zaluzec get chance to sit down and organise the files. So, please be patient. Many thanks.
Dear Em specialists, since a few months we are dealing with a stupid problem: every time we prepare samples for classical em investigation we sometimes have contrast and sometimes not. The procedure we uses is: 3% glutaraldehyde fixation (from 1 hour to overnight), wash in 0.1 M cacodylate (approx 30 min) buffer, osmiumtetroxide fixation (1 or 2% in caco for approx 1h), again a wash in caco (30 min), dehydration in a ethanol series (70, 90, 2x100 every step 30 min) then a 2X10min incubation in propylene oxide, an overnight impregnation in 1:1 epon (lx): propylene oxide and finally an embedding in lx at 60 degrees for 60 hours. After sectionning, the sections are contrasted for 2 to 3 min with 5 % uranylacetate in water and finally 1 min lead citrate. Every time we use exactly the same procedure but the contrast varies from perfect to absent. We have already tried to change the fixatives and contrasting solution , prepare fresh, change lot, change companie but nothing helped. Also we changed the incubation times but now ower inspiration is gone so ... who can help us solving this irritating problem? Thanks for your help. Annette bakker
Annette wrote: } since a few months we are dealing with a stupid problem: every time we } prepare samples for classical em investigation we sometimes have } contrast and sometimes not. The procedure we uses is: } 3% glutaraldehyde fixation (from 1 hour to overnight), wash in 0.1 M } cacodylate (approx 30 min) } buffer, osmiumtetroxide fixation (1 or 2% in caco for approx 1h), again a } wash in caco (30 min), dehydration in a ethanol series (70, 90, 2x100 } every step 30 min) then a 2X10min incubation in propylene oxide, an } overnight impregnation in 1:1 epon (lx): propylene oxide and finally an } embedding in lx at 60 degrees for 60 hours. After sectionning, the } sections are contrasted for 2 to 3 min with 5 % uranylacetate } in water and finally 1 min lead citrate. Every time we use exactly the } same procedure but the contrast varies } from perfect to absent. We have already tried to change the fixatives and } contrasting solution , prepare fresh, change lot, change companie but } nothing helped. Also we changed the incubation times but now ower } inspiration is gone so ... who can help us solving this irritating problem? } Thanks for your help. } Annette bakker
Hello Annette! I think we all have had more or less problems getting optimal contrast in our specimens but there are also the ugly precipitates which can terrify us. We poststain with UAc (2.5% ethanol solution) for 10 min or with 5% water solution of UAc for 20-30 min, followed by staining with fresh Pb-citrate for 2 min. I think your staining periods are too short. The staining results are also depending on what tissue you work with. UAc reacts prin- cipally with nucleic acids and proteins. Pb-citrate increases the general contrast of membranes and also stains nucleic acid, glycogen and proteins due to the presence of carboxyl and sulfhydryl groups. Good luck and feel free to contact me for further discussions. Sverker Enestr|m, M.D., Ph.D. Department of Pathology Link|ping university, Sweden
Kem Rogers asked about image enhancement for cytochemistry
Here is my two cents worth, As an ardent image processor enhancer, I have no apriori bias against using image enhancement. However, in cytochemistry if you are comparing two things, I think both images should be taken under the same conditions and processed equivalently. Kem mentions processing to remove background, and under the circumstances he outlines this seems reasonable. However, I think background substraction should be done with great care. One mans background could be another mans signal. If you have measured background under conditions where no signal is possible and then subtract it uniformly from all images that seems reasonable. However, I have had people ask me to remove this defect or that "small area of background staining". I think that begins to constitute scientific fraud. Thats my humble opinion.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
concerning your staining problem: 1) try doing an "en bloc" stain with 2-4% UA, after OsO4 and your buffer wash, and just before your dehydration step. simply immerse your tissue in UA for 30-90 min (keep at 4 degrees C) then proceed with dehydration 2) when staining your sections with UA and Pb citrate, staining times should be 10-15 min. in each stain
Annette, I agree with Dr. Sverker. The staining times are too short. I stain for 25 min in uranyl acetate and 15 min in Pb citrate. These times may seem excessive but we have never had a problem with contrast. Additiionally, more than one rinse is suggested by my sources (at least 3/15 min each) in rinse buffer to remove excess glutaraldehyde. This should eliminate any interaction between unreacted glutaraldehyde and OsO4 which will cause peppering and general visual noise in your sections.
Hope this is helpful!
Dirk W. Domaschko Marshall University Huntington, WV Domaschk-at-musom01.mu.wvnet.edu
In addition to the sticky from cellophane tape, thick sections which are notoriously difficult to keep in place can be stuck to grids using unpolymerized epoxy resin. Dip one side of the grid to the top of a drop of epoxy, use compressed air to blow away excess resin from the open spaces, adhere the section and cure the resin in the oven.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
I'm not allowed to give the name of my reference, but "word has it" that the pH of the lead citrate is very important: if it is } 12, there will be little staining, and it may in fact bleach the sections (and lots of ugly contamination if much {12). Something to be checked each time the lead citrate is made. Also, small, unobvious changes in pipette bore, or how the pipette is held can cause significant changes in drop size, therefore in amount or concentration of solution delivered. Phil Oshel U Illlinois Center for Electron Microscopy oshel-at-ux1.cso.uiuc.edu At 10:18 AM 1/18/95 +0100, Willem.Jacob wrote: } Dear Em specialists, } since a few months we are dealing with a stupid problem: every time we } prepare samples for classical em investigation we sometimes have } contrast and sometimes not. The procedure we uses is: } 3% glutaraldehyde fixation (from 1 hour to overnight), wash in 0.1 M } cacodylate (approx 30 min) } buffer, osmiumtetroxide fixation (1 or 2% in caco for approx 1h), again a } wash in caco (30 min), dehydration in a ethanol series (70, 90, 2x100 } every step 30 min) then a 2X10min incubation in propylene oxide, an } overnight impregnation in 1:1 epon (lx): propylene oxide and finally an } embedding in lx at 60 degrees for 60 hours. After sectionning, the } sections are contrasted for 2 to 3 min with 5 % uranylacetate } in water and finally 1 min lead citrate. Every time we use exactly the } same procedure but the contrast varies } from perfect to absent. We have already tried to change the fixatives and } contrasting solution , prepare fresh, change lot, change companie but } nothing helped. Also we changed the incubation times but now ower } inspiration is gone so ... who can help us solving this irritating problem? } Thanks for your help. } Annette bakker } } } } Phil Oshel oshel-at-ux1.cso.uiuc.edu Center for Electron Microscopy Univ. of Illinois (Urbana) (217) 244-3135
We operate 7 electron microscopes (4 scanners, 2 TEM's, 1 microprobe), optical photomicroscope, 2 ultramicrotomes, materials science specimen prep equipment, high vacuum coater, ion beam coater, darkroom, etc - pretty much the generic general EM lab. At present we use a paper-based (dog-eared diary) booking system for scheduling users on the equipment.
I know that there are many labs that use electronic booking systems. Is this worthwhile, or does it cause more problems than it solves? Nearly all our users have access to ethernetted PC's. Information on pros and cons, problems, solutions, which software etc would be most useful. Our LAN superviser is especially interested in being pointed in the right direction for sources of software that will do the job.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
I implimented this type of approach back around 1990, and have found it to be very useful. (My source code is very simple since it is designed for VT100 emulation across the ethernet, so I would not recommend it.) Apart from simple security issues such as making sure that authorized (safe) people only use the microscopes, the biggest advantage is speed. Noone has to spend hours collating the pages, and you do not have to worry about honesty -- how many people write down all the hours that they use? The major issue is not the software (trivial to write) but arranging electronic locks for the hardware such that they can only be used with an appropriate password (different for each user). We got an electronics technician to build a simple card for our TEM to control the beam. I would be interested in hearing of any more general solutions.
Reply... RE} EM Lab Scheduling Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
Here at Michigan our lab serves users from many departments (nuclear, materials, chemical , electrical, civil and mechanical engineering, physics, chemistry, geology, pediatric cardiology and pharmacy to mention a few) and they are spread out all over the city of Ann Arbor (about a five mile radius). Since there are University computer sites all over campus and many people have Macintosh computers or can at least get access to them and there is a campus-wide Appletalk network, we us a simple scheduling system based on a collection of Hypercard stacks. Each microscope, or other instrument that is schedulable, has a stack associated with it. These stack are stored on a server in our lab and are accessible from anywhere on campus. Each stack is a basic calendar with the days of the week listed in four hour block for the day time (~8-5) and 8 hour blocks in the evening and at night. Clicking on any one block of time allows the user to book that entire block or part of it. The user needs a user name and password and trained users are issued these ids that are stored within the stack. users can book up to 1 week in advance normally but I have the option of overriding that. When the stack reaches a certain size it is archived and the current data is copied to a new stack. It is not a really secure system but it works well for us. If anyone wants a copy of any of the stacks to modify for their own purposes, I will make them available on freebie.engin.umich.edu. The proviso is that any improvements should be made available to everyone, us especially!. OK? Just my 2 cents worth. Jfm.
I am interested in the discussion of light element analysis using a Kevex Quantum detector. From my discussions with Kevex, it seems that the software available has a hard time compensating for matrix effects when light elements are mixed with high atomic number elements. I was wondering how other people deal with this. Our system runs on RT11, so people who have TSX systems will have to bear with me. It appears that the answer is standards, standards, and more standards. Let's hear it... Randy Nessler
A couple of questions on CPD: 1. How important is it to use CO2 from a cylinder furnished with an ejector tube?
2.Can one use a conventional cylinder turned upside-down?
3.Am I right in assuming that cooling the CPD-chamber down to { 0 C, should condensate the CO2 to a liquid?
4.What grade (purity) should one use? I have always used "Medical Grade", obviously at a higher cost than "Technical Grade"
Discussion would be appreciated.Alan Hall Unit for Electron Microscopy University of Pretoria, Pretoria Tel +27-12-420-3297 South Africa Fax +27-12-420-3266
Could anyone please direct me to sources of Anopore disc filters or Flotronics filters for use in support of particulate samples for SEM viewing. A Canadian or US supplier would be helpful. Thanks Carolyn J. Emerson Biology Dept. Memorial University St. John's Newfoundland Canada cemerson-at-kean.ucs.mun.ca
I have someone interested in doing some reflection electron microscopy. Not being the surface type, I am unable to assist. If anyone out there is remotely interest in doing some REM please contact me directly for more details.
Thanks for your time.
Jeff --------------------------------------------------------- U U | | U U | Jeff Shield | U U | Department of Materials Science and Engineering | U U U U | University of Utah | U U U U | Salt Lake City, UT 84112 | U U U U | 801/581-3179 | UUUUU U | Fax: 801/581-4816 | U U | | U U --------------------------------------------------------- UUUUU
Of making many books there is no end, and much study wearies the body. -Eccl 12:12
AMRAY has been very good about supplying parts for our 1000A (still our workhorse 4-6 full days per week). Their telephone number is 800-225-1462. We've also had to replace some of these switches after some years.
} We are trying to repair the push-push type switches in the control } panel of our AMR 1000 SEM. The failure is mechanical rather than electronic. } The switches have "Master Specialties Co." writen on them and are in the } 12-4 series. Any line on where we can beg or buy at least two such switches } would be greatly appreciated. } Thanks in advance. } "For want of a nail...." } Tom Hanley } 706 568-2075 } thanly-at-uscn.bitnet
Jacob Jacob Bastacky, MD 1-116 Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Reply... RE} CD ROM image archive Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
While we are on the subject, has anyone seen the Pinnacle Micro drive for $1995 that is a double speed reader and also a CD-R drive? Sounds really nice!
You can read about reflection microscopy (which visualizes zones of cellular attachment to glass in epi-illuminated specimens by surface reflection interference) in chapter 5 in ELECTRONIC LIGHT MICROSCOPY, edited by David Shotton, Wiley-Liss Inc., 1993. Modern REM makes use of video-enhanced contrast technic, well described by David Shotton.
*************************************** Petra M Nederlof, Ph.D. Max-Planck-Institute for Biochemistry Dept. Structural Biology Am Klopferspitz 18 a D-82152 Martinsried (M=FCnchen) GERMANY
I have a Tektronix Phaser II SDX printer that I want to put on my campus network. I bought the Ethernet adaptor card but my network administrator is telling me it's not possible to put the printer on our network. We support Novell and TCP/IP. Has anyone put this printer on a network using either of these protocols? If so, can you give me any words of wisdom that might help me get the printer on the net?
Steven J. Eppell Center for Cardiovascular Biomaterials Case Western Reserve University
MICROBEAM ANALYSIS SOCIETY STUDENT SCHOLARSHIPS TO MAS-1995 (Breckenridge, CO) and IUMAS-1 (Sydney Australia)
All students (and faculty) involved in microanalysis-related research, should note a remarkable opportunity for travel to two forthcoming microanalysis conferences. The Microbeam Analysis society is offering student scholarships to the 29th Annual MAS meeting in Breckenridge CO (August 6-11, 1995). Student papers must be submitted, in standard MAS-MSA format to Dr. E. Etz, NIST, Division 837, Bldg. 222, room Q-113, Gaithersburg, MD 20899 by March 15, 1995. The best submitted papers will be awarded funds towards attending the annual meeting in Breckenridge. Any more information about the program can be obtained from Dr. Etz at etz-at-gapnet.nist.gov
What makes this scholarship offer extraordinary is that the best three papers given by student scholarship winners at the Breckenridge meeting will be awarded a minimum of $500 towards the cost of attending the 1st International Union of Microbeam Analysis Societies Conference in Sydney, Australia, February 5-9, 1996. These scholarships are only open to student members of MAS, and student application forms for MAS are available in past issues of Microbeam Analysis, the MAS journal. Student membership is a great bargain at $2.50, and doesn't require that the advisor be a MAS member - although if you aren't, I ask that you consider joining.
I will mail you more information with appropriate details about the meetings and the paper format etc., but if you have any immediate questions, please don't hesitate to contact me by email (dbw1-at-lehigh.edu).
we connercted our printer up for multiple users via the ethernet connection -first check to see that the installed tag is next to the ethernet port (if not you need a bourd installed) -connect to the ethernet port -load the phaser utilities, the network utilities, and the drivers -there are a few entries (words/code #'s/and commands) needed to load it all and get it going -if all else fails call Tektronix
good luck Mike
On Mon, 23 Jan 1995, Steven J. Eppell wrote:
} I have a Tektronix Phaser II SDX printer that I want to put } on my campus network. I bought the Ethernet adaptor card but } my network administrator is telling me it's not possible to } put the printer on our network. We support Novell and TCP/IP. } Has anyone put this printer on a network using either of } these protocols? If so, can you give me any words of } wisdom that might help me get the printer on the net? } } Steven J. Eppell } Center for Cardiovascular Biomaterials } Case Western Reserve University } } sje-at-po.cwru.edu }
Steven} I have a Tektronix Phaser II SDX printer that I want to Steven} put on my campus network. I bought the Ethernet adaptor Steven} card but my network administrator is telling me it's not Steven} possible to put the printer on our network. We support Steven} Novell and TCP/IP. Has anyone put this printer on a Steven} network using either of these protocols? If so, can you Steven} give me any words of wisdom that might help me get the Steven} printer on the net?
Steven} Steven J. Eppell Center for Cardiovascular Biomaterials Steven} Case Western Reserve University
Steven} sje-at-po.cwru.edu
If you are trying to print from a Unix workstation you can use the standard BSD print spooling mechanism to print to a network node (the IP address assigned to the Ethernet adaptor. If this is a Novell network and you need to print from a Sun Unix workstation you can use a package from Puzzle Systems called SoftNet Utilities that allows bidirectional printing.
-- Kevin Burton Noran Instruments voice: (608) 831-6511 x317 2551 West Beltline Highway, Room 532 FAX: (608) 836-7224 Middleton, WI 53562 email: kburton-at-noran.com
All subscribers Please remember to change your default address of "microscopy" to
Microscopy-at-aaem.amc.anl.gov
ANLEMC is dead, and mail is aliased to the new site.
Unfortunately the alias does not always work so some messages to microscopy do not always come through.
--------------------------- As for your Tek IISDX....
You should likely talk to Tektronics people on your network problem. I have a IISDX which is on an Appletalk/Ethernet connection and works fine. There are special drivers which you must load for using the printer on Mac, PC, or Sun workstations. You should have gotten these with your hardware. You could also run a direct line between the printer and your workstation.
On the Mac side you simply, plug-in and run. Virtually no setup time is required, except to copy the drivers from the disks to the system folder and connect the printer to the net. Then simply use the "choozer" to locate the printer on the appropriate zone on your networks. (This is the network connection I use)
On the PC side you will have to install the Windows drivers and then Mount the printer as a network printer. However, I cannot help with Novell nets.. How do you normally connect network printers? You must have laserprinters on your Novell network and I would guess that the procedure would be similiar. My version of the IISDX is NOT IP aware (it's about 2 years old) so TCP/IP is not an option here.
Just a warning. Putting a dye sub on a network can also be expensive if you have users that are not careful. People often forget that they were using a dye-sub printer and when they want to print their next manuscript/email note sometimes send it to a printer assuming they are still connected to a standard text printer, without checking to see that the have "de-selected" the dye sub. Hope you have some ideas on how to limit access otherwise you may end up having lots of expensive text printed out on your dye sub.
I use the IISDX printer for literally all my image hardcopy now. Haven't made a print from TEM negatives in nearly 2 years. Of course, some of my instruments are entirely digital (VG HB603Z) and hence there is no film.
Message-Id: {9501232034.AA26017-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Light element EDS Orig-Author: {randy nessler {rnessler-at-emiris.iaf.uiowa.edu} }:ddn:wpafb -----------------------------------------------------------
I am interested in the discussion of light element analysis using a Kevex Quantum detector. From my discussions with Kevex, it seems that the software available has a hard time compensating for matrix effects when light elements are mixed with high atomic number elements. I was wondering how other people deal with this. Our system runs on RT11, so people who have TSX systems will have to bear with me. It appears that the answer is standards, standards, and more standards. Let's hear it... Randy Nessler
An eductor tube ie a syphon tube (not ejector) is usual to get the liquid phase of co2 out of the tube. Ris at Univ Wisc used a small cylinder upside down, not very pratical for a large cylinder. I don't think it practical to get the cylidar down to a low enough temp to get liquid out, I forget the liquification temperature but it must be close to -70 C ie close to dry ice temp? Quality is important, I have had cylinders that had water mixed with the co2.... bad scene!! I order extra dry co2 (I believe, its written down where I order, not here) MOST important! be sure that the syphon/eductor tube is really there! Test by letting gas out at a good rate, the valve should start to frost up within 5-10 seconds at at a reasonable flow. about 20 percent of cylinders have broken or missing eductor tubes! so beware. A 'condenser valve' is available, I think from Fullam? (mine is ~12 years old) this will bleed some c02 out while cooling an internal cold finger that will recondense gaseous co2 passing through the valve to the cpd back to liquid... provided the room temp is not too high and the cylinder is not too empty etc. Also only about 30 of the 50 pounds of co2 can be gotteon out as liquid even with a condensing valve. Cold water around the chamber also helps. Hope this helpes people. alan pooley marine sci sem lab rutgers univ
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu Time: 4:46 PM
Date:1/23/95 URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html NC EMAL
Hi there, we are in the process of converting our JEOL 4000EX from freon to SF6 and want to know where we can get the said gas at a reasonable price. At the moment it looks like we are looking at between $1000 and $1700 for a single charge and so I would appreciate it if anyone could give me pointers for a source with a more reasonable cost Please reply direct to me and I will summarize to the list if there is sufficent interest. Thanks John Mansfield.
Ta muchly - hooked up and ready to go Brendon J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-9-380-2739 fax 61-9-380-1087
we're wanting to get back into the lucifer yellow business after a few years - filling cells in slices, observing the fills with fluorescence, then immunostaining with antibody against LY and through to DAB. In the past, we used a gift antibody. Are there commercial Ab's out there that also do a good job? I have heard that there can be some loss of fine detail in the immuno procedure, and want to know if anyone has strong preferences as to LY antisera.
------------------------------------------------------------------ |Glenn Holm *mime mail ok* Internet:karuzis-at-wccf.mit.edu | |M.I.T Dept. of Brain + Cog. Sci. This VAX doesn't do NeXTmail | |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | ------------------------------------------------------------------
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John In my recent experience, SF6 is available from all of the major industrial gas suppliers (ie Selox) for about the same price...~$700 per 100 lb. That was enough for about 3 charges on our JEM 1210. Your 4000 must have a BIG tank. It took about 6 weeks for our SF6 to arrive. Obviously there is some primary supplier, but I don't know where. Good luck!
buddy
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
A few months ago, I put a message on the board looking for a Freeze Fracture machine, didn't get any good leads, so I thought it was time for another try:
We would like to obtain a freeze fracture machine in good working order (one of the Balzers or Cressington machines would do fine). We have money to pay for the machine, though not enough for a new one (sound familiar!). If you or someone you know would be interested in selling a machine to a good home please let me know
Thanks. ---------------------------------------------------- Simon Watkins Ph.D Director Structural Biology Imaging Center Scaife 840 University of Pittsburgh Pittsburgh PA 15261
The price for SF6 depends greatly on the purity. Commercial purity (99.7%) is generally available around $650-700 for a cylinder (115lb capacity). Instrument grade (99.99%) is near $1100 per cylinder. Our prices are from AGA (about a year ago), although I don't think you'll find significant price differences from other vendors.
The microscope manufacturers recommend instrument grade (of course!), although we successfully ran our H9000NAR on commercial grade.
Cheers, Henk
} } Hi there, } we are in the process of converting our JEOL 4000EX from freon to SF6 and } want to know where we can get the said gas at a reasonable price. At the } moment it looks like we are looking at between $1000 and $1700 for a single } charge and so I would appreciate it if anyone could give me pointers for a } source with a more reasonable cost } Please reply direct to me and I will summarize to the list if there is } sufficent interest. } Thanks } John Mansfield.
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- Research Law #1 If observations disagree with theory, the observations are obviously in error.
I surveyed sources of freeze drying units last year when finishing up my book on "Vacuum Methods in Electron Microscopy". At that time, these devices were available from: Agar Scientific (UK) Fx:0279-815106 Bal-Tech Fx: 508-374-7070 Denton Vacuum Fx: 609-424-0395 Edwards High Vacuum Fx: 505-658-7969 Electron Microscopy Sciences Fx: 215-646-8931 Emitech Fx: 713-893-8443 Energy Beam Sciences Fx;413-789-2786 Fisons FX: 415-961-8656 GeneVac (UK) FX: 0473-461176 Microfield (UK) FX:0865-821459 VG Microtech (UK) FX: 0825-768343 I don't know about the degree of automation offered, but perhaps this will give you a start on sorting things out. Good Luck!
--------------------------------------
------------------------------------------------------------------------ | David O'Neil tel: (902) 426-8258 | | National Research Council of Canada fax: (902) 426-9413 | | Institute for Marine Biosciences _____ _____ | | 1411 Oxford St. | | __/\__ | | | | Halifax, Nova Scotia B3H 3Z1 | | __/\\ //\__ | | | | Canada | | \ \\ // / | | | | | | /___ ___\ | | | | email: oneild-at-imb.lan.nrc.ca | | /__ __\ | | | | | | || | | | | |_____| |_____| | ------------------------------------------------------------------------
------------------ RFC822 Header Follows ------------------ Received: by mse.engin.umich.edu with SMTP;24 Jan 1995 12:59:48 -0400 Received: by truelies.rs.itd.umich.edu (8.6.9/2.2) with X.500 id MAA14718; Tue, 24 Jan 1995 12:58:43 -0500 with SMTP id MAA14681; Tue, 24 Jan 1995 12:58:39 -0500 To: Microscopy-at-AAEM.AMC.ANL.GOV
This is a follow-on to the 'pre-announcement' that I posted several weeks ago. The position is now real.
Materials Science and Engineering Laboratory National Institute of Standards and Technology
We are presently expanding our capabilities within the Materials Science and Engineering Laboratory at NIST in the area of HREM, and have an opening for a highly qualified Ph.D. scientist. Candidates must have extensive hands-on experience in high resolution imaging, be well versed in imaging and diffraction theory, and have exper- ience in image simulation. Experience in PEELS and EDS is essential, as is some background in materials science. Strong interest in development and implementation of new methodology is necessary. The position will include extensive work with other staff members of MSEL. For permanent employment, US citizenship is required.
Interested parties should submit a curriculum vitae and the names of three references by MARCH 15, 1995 to:
Dr. F.W. Gayle NIST Building 223, Room B164 Gaithersburg, MD 20899
Ed, I tried to respond to your request for print processor information, but my attempt bounced back with "host unknown" message. Please contact me privately and send e-mail address and I'll try again.
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Good day all, Thanks to all the contributors who posted (and are still posting) responses to my question on electronic lab work scheduling. Shall try to post a summary of the responses.
Another query: We need to put together a cooling stage for freeze drying of small samples. One way of doing this could be by using Peltier-effect cooling. These devices are used in various pieces of equipment but I have not been able to find someone who makes them. Could anyone help by supplying references to manufacturers of Peltier-effect devices? Thanks
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
I was looking for such a unit for years. I am currently using unit which was build in our workshop for 10 years ago and still going strong. The unit have rotary and turbo-pumps. For cooling Neslabs cryocool and liquid nitrogen are used. Temperature of specimens down to -150C and condenser, down to -180C. Using a data acquisition card from Strawberry Tree + Mac I can use controll and register temperatures at 4 different sites and register and display vacuum. Freeze-drier is used for both SEM-preparations and Low Temperature Vacuum Embedding. I will present the latest results from this system at SEM-meeting in Houston, Texas in May 1995. Costs of total unit are about USD 10 000 INCLUDING pumps, acq-card (not crypcool).
Regards
Romuald
------------------------- Romuald Wroblewski, Ph.D. Associate Professor Department of Pathology Karolinska Institute voice/fax:+46-8-7293597 -------------------------
I thought I would poll the EDS, WDS, EELS and other users about the proper disclaimer to place in reports issued internally or externally.
Most of our analysis is concerned with mineral fillers and those elements heavier than neon dispersed in an organic matrix. In some cases we will examine the organic matrix or we ash the organic matrix and examine the ash. The ash is either pressed on to a carbon block or on to a conductive double-sided tape. We believe that a realistic blanket statement of detectable for semi-quantification mode is "between neon to neptunium if present in concetrations of approximately 0.5%" but we feel a need to add some weasel words about the sample.
The question: Which best describes the limitations of our analysis by our samples? 0.5% by weight or 0.5% by volume, or is there a better phrase? Or are we just fooling ourselves?
I thought I would poll the EDS, WDS, EELS and other users about the proper disclaimer to place in reports issued internally or externally.
Most of our analysis is concerned with mineral fillers and those elements heavier than neon dispersed in an organic matrix. In some cases we will examine the organic matrix or we ash the organic matrix and examine the ash. The ash is either pressed on to a carbon block or on to a conductive double-sided tape. We believe that a realistic blanket statement of detectable for semi-quantification mode is "between neon to neptunium if present in concetrations of approximately 0.5%" but we feel a need to add some weasel words about the sample.
The question: Which best describes the limitations of our analysis by our samples? 0.5% by weight or 0.5% by volume, or is there a better phrase? Or are we just fooling ourselves?
Just a note. It is clearly inappropriate to use the Microscopy Listserver to send out Test messages to see if your Email server is running. There are somewhere over 2000 people receiving these messages please be intelligent. If you are having an Email problem then send a message to only one person. If you are really in a bind then just send a message to me and I will respond when I get a chance, but not to the world!!
Nestor Your Friendly (& Lately Tired) Neighborhood SysOp
I am planning to teach a short course in biological SEM for novices and need to find a textbook for the students. I am considering Postek et al., 1980 published by Ladd. Would any of you out there recommend anything more recent that would be appropriate for grad students, staff, faculty, post-docs. Any input willl be appreciated.
Although my course is intended for local consumption, iot would be open to anyone, so if there are interested parties wanting to spend a week in Florida in May, just E- mail me at the address below
I posted this note yesterday on several newsgroups, but just found out about this listserver today (1-25-95). I am looking for some help...
My group is responsible for examining uranium-metal spent nuclear fuel (N-Reactor, Single-Pass Reactors) which has degraded over time while in water storage at the Hanford, Washington site. There is a possibility that some of the uranium may have reacted with water to form uranium hydride which still may be present in some of the fuel. One of our tasks is to determine if this scenario is real. Thus, we will be examining several fuel elements to look for evidence of hydrides. We have (and will use) optical microscopy and electron microscopy (AFM, SEM, TEM) in our efforts.
Because the fuel is highly radioactive, and has a potential for pyrophoric reactions, our first efforts will be to cut, section, polish, and optically examine the fuel in a radiation hot cell using an argon cover gas blanket over all operations. However, when we get to the point of specifying etchants or etching techniques to examine the uranium, no one is sure how best to bring out the features of uranium hydride in uranium metal. We will try cathodic etching first, largely because this is what has been traditionally done to examine uranium at our lab. However, we aren't sure what to expect from the hydride (if it exists). Our literature searches have drawn a blank with regard to microscopy techniques for uranium hydrides.
So, I would like to ask, has anyone had experience optically examining uranium hydrides? If so, could you recommend a procedure? We also have the possibility of hiring a consultant if the right person or lab anywhere in the world) is identified to assist us.
I have a question for those people involved in light microscopic immunolabelling and trying to quantify the amount of stainig. Can anyone do it reliably?? We have someone in the lab who wants to determine if the staining obtained with 6 different antibodies co-localize within the tissue. They are staining paraffin-embedded sections and are staining one antibody/section. They want to know if there is a way to do this using an imaging system of some sort. IMHO I don't thinnk it can be done easily. If anyone has any trhoughts/ideas they would be GREATLY APPRECIATED.
Thanks Mark Elliott, PhD Pulmonary Research Lab UBC Vancouver Canada
Quantitative fluorescence microscopy can be tricky. Please refer to the chapter by Jesse E. Sisken (Fluorescence Standards) in Methods in Cell Biology, vol. 30, edited by L.Taylor and Y. Wang (1989; Academic Press) for recommendations.
************************************************************************* M.V. Parthasarathy Professor of Plant Biology Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14850 Telephone: (607) 255-1734 Fax: (607) 255-5407 E-mail: mvp2-at-cornell.edu ***********************************************************************
Try using silver paint, we use it & it seems to be working well. -Mike
On Wed, 25 Jan 1995 tsu-at-cae.wisc.edu wrote:
} } Hi, microscopist, } } Does anyone know if there's any conductive glue by which we can attach } our TEM samples to Cu/Au grids? M-bond is what I currently use and has } been suspected to cause some shifting of images especially for tiny } samples. Any information or comment will be appreciated. } } I-Fei Tsu } ASC-MRG } UW-Madison } }
Does anyone know if there's any conductive glue by which we can attach our TEM samples to Cu/Au grids? M-bond is what I currently use and has been suspected to cause some shifting of images especially for tiny samples. Any information or comment will be appreciated.
I am trying to prepare TEM thin foils from Mo-47.5Re (at.%) using a twin-jet electropolisher, without success. I have tried cold sulfuric acid/methanol solutions and perchloric acid/methanol solutions. These produce a pebbly surface with minimal electron transparent areas.
Does anyone out there have experience producing TEM thin foils from Mo-Re? I would greatly appreciate any suggestions! _ _ O-O J (a sideways smiley) Thanks, Dan Schwartz ~
I spoke to Bernie Kestel at Argonne National Lab and he gave the following solution that has worked well for him:
Material: Mo 30a/o Re Equipment: South Bay Technology Model 550 Single Vertical-Jet ElectroPolisher Electrolyte: 12% Sulphuric Acid 83% Methanol 5% Butyl Cellosolve Temperature: -50C Voltage: 190V Current: 50-75 ma Pump Speed: Slow
Since the work was done with a SBT Single Jet System, he suggested that you change the temperature to perhaps -40C and use a somewhat lower voltage for a twin jet system.
Of course, my solution would be to buy a South Bay unit, but I do have a vested interest in that! We also publish a bibliography of technical reports on sample preparation and distribute copies of the papers free of charge. Most of the reports deal with TEM sample preparation - primarily electropolishing, dimpling and Tripod Polishing.
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
Message-Id: {m0rXYnF-0000zMC-at-pegasus.cc.ucf.edu} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Electropolishing Mo-Re Orig-Author: {schwartz-at-mrvx03.mdc.com}:ddn:wpafb ----------------------------------------------------------- My Fellow Microscopists:
I am trying to prepare TEM thin foils from Mo-47.5Re (at.%) using a twin-jet electropolisher, without success. I have tried cold sulfuric acid/methanol solutions and perchloric acid/methanol solutions. These produce a pebbly surface with minimal electron transparent areas.
Does anyone out there have experience producing TEM thin foils from Mo-Re? I would greatly appreciate any suggestions! _ _ O-O J (a sideways smiley) Thanks, Dan Schwartz ~
This may be a simple minded question, but I could use some advice. I just mixed some Epon Araldite type embedding plastic and it got really dark in color after adding the BDMA. I was using an Epon 812 substitute, DDSA, Araldite 502, and BDMA. It is a slightly different recipe than we have used before and our plastic is usually a pleasant golden color, this stuff was like dark caramel in color. The color came from the interaction of the BDMA and DDSA, or at least that is the way it looked after mixing the BDMA with the other components individually.
Does anyone have a suggestion to explain the dark color and any free advice about this phenomenon in general.
In a related question, can anyone say what the useful shelf lives of the various embedding components might be. Some of ours are older, someone buys a lot and never uses it up, should we toss everything and start over or can we hold on and use what we have?
Thanks for your help.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95060 emlab-at-ucsco.ucsc.edu (408) 459-2477
I DON'T THINK THE AGE OF YOUR RESIN COMPONENTS HAS ANY INFLUENCE ON THE DARK COLOR. OUR RESIN BATCHES ARE TURNED-OVER RAPIDLY, AND WE HAVE NOTICED A DARK COLOR IN OUR SPURR IN COMPONENTS THAT WERE RECENTLY ACQUIRED. WE CALLED ONE OF THE SUPPLY COMPANIES AND WERE TOLD THAT AN ALTERED PH IN ONE OF THE COMPONENTS WAS RESPONSIBLE. SINCE OUR SPURR RECIPE AND YOUR RESIN RECIPE DO NOT HAVE ANY COMPONENTS IN COMMON, PERHAPS THIS IS A GENERAL RESIN PROBLEM. ANY COMMENTS FROM RESIN ENGINEERS?
MARGE
Marge Hukee EM Core Facility hukee.margaret-at-mayo.edu Mayo Foundation (507) 284-3148 ---------------------------------------------------------------------------- --
About to enjoy a break.....please unsubscribe, and thanks.
Ron Oldfield School of Biological Sciences Macquarie University NSW Australia 2109 email: roldfiel-at-rna.bio.mq.edu.au phone: (612) 850 8173 fax: (612) 850 8116
Mats Karlsson (at Dept. of Pathology, Orebro Medical Center Hospital, Orebro, Sweden) et al. have recently described the methodological approach to quantify immunostained objects in histological sections by computerized image analysis in: Pathology, Res. and Pract. 1995 (in press), using frozen sections. Intra- and interindividual variations were less than 5% and the correlation of reproducibility was high. If you are interested, contact me directly.
================================================================== Sverker Enestrom M.D., Ph.D. Department of Pathology University of Linkoping, Sweden Phone: +46 13 22 15 20 Fax: +46 13 13 22 57 ==================================================================
Does anyone has experience how to get channeling patterns on JEOL Jem 2000FX electron microscope equipped with ASEA20 in STEM/BEI mode? Please some hints, advices.
Thank You.
Peter Makroczy Technical University of Kosice Dept.of Materials Science Park Komenskeho 11 043 85 Kosice Slovak republic
Message-Id: {9501271212.AA08568-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Electropolishing Mo-Re Orig-Author: {schwartz-at-mrvx03.mdc.com}:ddn:wpafb ----------------------------------------------------------- My Fellow Microscopists:
I am trying to prepare TEM thin foils from Mo-47.5Re (at.%) using a twin-jet electropolisher, without success. I have tried cold sulfuric acid/methanol solutions and perchloric acid/methanol solutions. These produce a pebbly surface with minimal electron transparent areas.
Does anyone out there have experience producing TEM thin foils from Mo-Re? I would greatly appreciate any suggestions! _ _ O-O J (a sideways smiley) Thanks, Dan Schwartz ~
Does anyone has experience how to get channeling pattern on JEOL Jem 2000FX electron microscope equipped with ASEA 20 in STEM/BEI mode. Please some hints, advices.
Thank You.
Peter Makroczy Technical University of Kosice Dept. of Materials Science Park Komenskeho 11 043 85 Kosice Slovak republic
Way back when, I was using EPON-ARALDITE resin components that were at least 10 years old at the time. No problems with using these. I do not recall having any problems with a dark color. What % of BDMA are you using? I used 2%(v/v). The only problem I can forsee in using a dark batch of resin would be that your background of your sections will not be as light as they should be. Best of luck.
It is hard to understand why people are still attempting to 'quantitate' imunohistochical reactions which rely on a final step in which the 'quantity' of stain is not related to the amount of antigen present. In order to quantitate anything there has to be a quantitative relationship which can be measured relating what is seen in the section to the amount of material detected. In other words you cannot quantiate the standard immunoperoxidase techniques used in immunocytochemistry. This does not negate their importance, but means that one should not attempt to do image analysis on this material.
For a complete discussion of these limitations see the bottom of page 6 in the Gareth Griffiths book on "Fine Structure Immuno-cytochemistry, Springer-Verlag, 1993, ISBN 3-540-54805-X. The following is a quote from this page "..., Fig 1 shows a striking example of a little appreciated fact, namely, how the immunoperoxidase method can often give results that, though qualitatively correct, may be quantitatively misleading. Most people who have worked extensively with any of the HRP techniques become aware of these problems and may be frustrated by the presence of many variables that cannot, even empirically, be completely controlled. Even if only qualitative data are required, it is very difficult in practice to find the right compromise between acceptable fine structure preservation and specific labelling. This does not belittle the historical importance of the immunoperoxidase techniques..."
Herbert K. Hagler, Ph.D. Microscopy and Imaging Service Center Dept of Pathology, UT Southwestern Medical Center 5323 Harry Hines Blvd., Dallas, TX 75235-9072 phone (214) 648-3890 fax (214) 648-3925
please, unsubscribe ------------------------------------------- Giovanni Valdre' Dipartimento Di Scienze Mineralogiche Piazza di Porta S. Donato 1 I-40126 Bologna, Italy Tel. +39 51 243556 FAX +39 51 243336 email gvaldre-at-geomin.unibo.it -------------------------------------------
Please send copies of Temhelp letters or questions to me. We perform all types of EM analyses and can probably help with a variety of techniques/comments.
Thank you very much.
Regards,
Mark W. Rigler, Ph.D. Materials Analytical Services Norcross, Georgia 1800-421-8451
Many different epoxies from various manufacturers, with widely varying ages have all given me exceptionally dark pot mixtures from time to time. There are only two things that I've been able to narrow it down to. One is accelerator, BDMA, DMAE or DMP-30 that has been opened too long. Water contamination has been suggested by mfrs. I now toss the accelerator when a bottle of epoxy (Epon, DER 736, etc.) is emptied. Some suppliers are worse than others. We quit dealing with Polysciences entirely, because of irregularities in the final blocks. (color is minor, it's when things won't get hard, and kits arrive missing components that things are serious). The other correlation on block color is delay in mixing. If you delay, you get coloration. Delay too long, or mix with a stir bar so slowly that you get layer formation, and the upper layer will get very dark and form lumps. Just my $.02.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
looking for feedback (mostly negative) from users - I have demo'd the systems and have a few "satisfied customer" refs from sales and of course each has annon. "horror stories" about the competition Hi everyone
I am in the process on deciding on a purchase of cryo-fixation and ultramicrotomy equipment and would appreciate some feedback from users -
I have demo'd both the Reichert Ultracut S and the RMC 7000 with their cryo systems and they seem pretty comparable, each having its own plusses and minuses -
This instrument will be used for both teaching and research - also it will msot likey be a very long time before we purchase another one
I would appreciate any positive and/or NEGATIVE comments you might have on these instruments - particularly your experience regarding reliability and service and support after the sales (I will not be taking out a service contract)
We are also contemplating the purchase of a jet freezer from either Bal-Tec or RMC (they bought out the Life-cell system) & would appreciate any comments from users about those instruments as well
Responses which are sent directly to me (as opposed to those posted to the list) will remain confidential
Thanks
Marcelle Gillott, Ph.D. Director, EM Laboratory University of Wisconsin-Milwaukee
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 181-187.
In situ analysis of microbial consortia in activated sludge using fluorescently-labelled, rRNA-targeted oligonucleotide probes and scanning confocal laser microscopy
M. Wagner, B. Assmus, A. Hartmann, P. Hutzler & R. Amann Lehrstuhl fur Mikrobiologie, Technische Universitat Munchen, Arcisstrasse 21, D-80290 Munchen, Germany
SUMMARY
Activated sludge flocs are complex consortia of various micro- organisms. The community structures of samples taken from municipal sewage treatment plants were characterized using fluorescently labelled, 16S and 23S rRNA-targeted oligonucleotide probes in combination with confocal scanning laser microscopy (CSLM). In comparison with conventional epifluorescence microscopy, CSLM considerably improved the capability to visualize directly the spatial distribution of defined bacterial populations inside the sludge flocs. Analyses could be performed at high resolution undisturbed by problems such as autofluorescence or limited spatial resolution in thick samples. In addition, CSLM was used to analyse some structural properties of paraformaldehyde-fixed activated sludge flocs, such as floc size and homogeneity. Typical floc sizes were found to be in the range between 5 and 50 micrometre. Whereas most of the flocs were completely colonized by bacteria, there were also examples of flocs containing gas bubbles or particles in the interior.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 188-194.
Scanning interference and confocal microscopy
R. Juskaitis & T. Wilson, Department of Engineering Science, University of Oxford, Parks Road, Oxford, OX1 3PJ, U.K.
SUMMARY
The form of the interference term image in scanning confocal and scanning conventional interference microscopes is identical in all respects including optical sectioning. This observation is used to obtain confocal images and surface profiles from conventional scanning interference microscope images.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 195-203.
Time- and wavelength-resolved spectroscopy in two-photon excited fluorescence microscopy
S. Andersson-Engels, I. Rokahr & J. Carlsson Department of Physics, Lund Institute of Technology, PO Box 118, S-221 00 Lund, Sweden
SUMMARY
Two-photon excited fluorescence spectroscopy has been performed at a microscopic scale in combination with normal, white light microscopy. This gave simultaneously a spectral resolution of 20nm and a temporal resolution of 20ps, from a volume element less than 5 micrometre in all three dimensions. The sample was excited with light from a continuously mode- locked Ti:sapphire laser that was focused on the sample in a fluorescence microscope. A polychromator and streak-camera were used for detection. The method has been used on tissue, plant and paper samples. It has also been demonstrated how substances naturally occurring in the samples can be identified from their spectroscopic properties and the spatial distribution of these substances can be observed.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 204-210.
Intracellular localization of the antitumour drug adriamycin in living cultured cells: a confocal microscopy study
S. Meschini, A. Molinari, A. Calcabrini, G. Citro & G. Arancia Department of Ultrastructures, Istituto Superiore di Sanita, Viale Regina Elena 299, 00161 Rome, Italy
SUMMARY
The intracellular distribution of the anthracyclinic antibiotic adriamycin in living cultured cells has been investigated by confocal microscopy. In human melanoma cells (M14), adriamycin was localized inside the nuclei. When adriamycin-treated M14 cells were allowed to recover in a drug-free medium, a complete efflux of the drug from the nucleus was revealed. In recovered cells, a weakly fluorescent signal was observed in the perinuclear region. When M14 cells were recovered in a medium containing colcemid, a microtubule depolymerizing agent, the drug transport from the nucleus to the cell periphery appeared to be inhibited, suggesting that the microtubule network is strongly involved in drug transport mechanisms. In multidrug- resistant (MDR) cells the intracellular location of adriamycin was shown to be noticeably different from that of the parental wild-type cells. In particular, in resistant human breast carcinoma cells (MCF-7), adriamycin appeared to be exclusively located within the cytoplasm, whereas the nuclei were shown to be completely negative. When adriamycin treatment was performed in association with MDR revertants, such as Lonidamine (inhibitor of the energy metabolism) or verapamil (inhibitor of the P-glycoprotein efflux pump), a marked enhancement of the cytoplasmic signal was observed in resistant cells. Under these conditions, adriamycin appeared concentrated in the perinuclear region, but the nuclei were still negative. Confocal microscopy proved to be a useful method for the study of the intracellular transport of fluorescent substances, such as anthracyclinic antibiotics, and for the investigation of the multidrug resistance phenomenon in tumour cells.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 211-221.
A versatile tilting device for fluorescence microscopes
J. Bradl, M. Hausmann, B. Schneider, B. Rinke & C. Cremer Institut fur Angewandte Physik, Albert-Uberle Strasse 3-5, D-69120 Heidelberg, Germany
SUMMARY
A tilting device for biological specimens (rotation angle up to 2 pi) especially fluorescence-labelled cell nuclei, was developed. It consists of a quartz glass capillary and a mounting adaptor for the microscope stage. The applicability of the device was tested for several epifluorescence and confocal scanning laser microscopes. The axis of rotation is perpendicular to the optical axis of the microscope. The capillary can be tilted around its axis at any desired angle or in equiangular steps. This can be done manually or by remote control using a stepping motor. The three-dimensional (3-D) image-forming properties of the capillary system were experimentally examined using an inverse confocal scanning laser microscope. The results were compared with measurements obtained from the same microscope with the standard stage for plane slides with cover glasses. The measured point spread function suggested that, in spite of aberration effects, the optical arrangement used allows a gain in the 3-D resolution by tilting the object. A low-cost, fully-automated 3-D imaging system was built on the basis of a conventional epifluorescence microscope with a cooled black-and-white CCD camera. The system was operated by a personal computer. The online visualization ('movie') of rotating objects indicates the feasibility of the system.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 222-225.
P. E. Hanninen, E. Soini & S. W. Hell Department of Medical Physics, University of Turku, Center for Biotechnology, Tykistokatu 6, FIN-20521 Turku, Finland
SUMMARY
Two-photon excitation fluorescence imaging is feasible with continuous wave lasers. Images of biological specimens are obtained by employing photon counting in conjunction with an increasing recording time. The approach allows two-photon three-dimensional imaging of fluorescently-labelled specimens with inexpensive lasers.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 226-230.
Refractive-index-induced aberrations in two-photon confocal fluorescence microscopy
H. Jacobsen, P. E. Hanninen, E. Soini & S. W. Hell Department of Medical Physics, University of Turku, Center for Biotechnology, Tykistokatu 6, FIN-20521 Turku, Finland
SUMMARY
The effect of refractive index mismatch on the image quality in two-photon confocal fluorescence microscopy is investigated by experiment and numerical calculations. The results show a strong decrease in the image brightness using high-aperture objectives when the image plane is moved deeper into the sample. When exciting at 740nm and recording the fluorescence around 460nm in a glycerol-mounted sample using a lens of a numerical aperture of 1.4 (oil immersion), a 25% decrease in the intensity is observed at a depth of 9 micrometre. In an aqueous sample, the same decrease is observed at a depth of 3 micrometre. By reducing the numerical aperture to 1.0, the intensity decrease can be avoided at the expense of the overall resolution and signal intensity. The experiments are compared with the predictions of a theory that takes into account the vectorial character of light and the refraction of the wavefronts according to Fermat's principle. Advice is given concerning how the effects can be taken into account in practice.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 231-237.
The tetrahedral tip as a probe for scanning near-field optical microscopy at 30nm resolution
U. C. Fischer, J. Koglin & H. Fuchs Westfalische Wilhelms Universitat, Physikalisches Institut, Wilhelm-Klemm-Strasse 10, 48149 Munster, Germany
SUMMARY
The tetrahedral tip is introduced as a new type of probe for scanning near-field optical microscopy (SNOM). Probe fabrication, its integration into a scheme of an inverted photon scanning tunnelling microscope and imaging at 30nm resolution are shown. A purely optical signal is used for feedback control of the distance of the scanning tip to the sample, thus avoiding a convolution of the SNOM image with other simultaneous imaging modes such as force microscopy. The advantages of this probe seem to be a very high efficiency and its potential for SNOM at high lateral resolution below 30nm.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 238-244.
Studies of porphyrin containing specimens using an optical spectrometer connected to a confocal scanning laser microscope
O. Trepte, I. Rokahr, S. Andersson-Engels & K. Carlsson Physics IV, The Royal Institute of Tech, S-100 44 Stockholm, Sweden
SUMMARY
A spectrometer has been developed for use with a confocal scanning laser microscope. With this unit, spectral information from a single point or a user-defined region within the microscope specimen can be recorded. A glass prism is used to disperse the spectral components of the recorded light over a linear CCD photodiode array with 256 elements. A regulated cooling unit keeps the detector at 277K, thereby allowing integration times of up to 60s. The spectral resolving power ranges from 350 at 400nm to 100 at 700nm. Since the entrance aperture of the spectrometer has the same size as the detector pinhole used during normal confocal scanning, the three-dimensional spatial resolution is equivalent to that of normal confocal scanning. Light from the specimen is deflected to the spectrometer by a solenoid controlled mirror, allowing fast and easy switching between normal confocal scanning and spectrometer readings. With this equipment, studies of rodent liver specimens containing porphyrins have been made. The subcellular localization is of interest for the mechanisms of photodynamic therapy (PDT) of malignant tumours. Spectroscopic detection is necessary to distinguish the porphyrin signal from other fluorescent components in the specimen. Two different substances were administered to the tissue, Photofrin, a haematoporphyrin derivative (HPD) and delta-amino levulinic acid (ALA), a precursor to photoporphyrin IX and haem in the haem cycle. Both are substances under clinical trials for PDT of malignant tumours. Following administration of these compounds to the tissue, the potent photosensitizer and fluorescent compound photofrin, or protoporphyrin IX, respectively, is accumulated. For our study Wistar/Furth rats were injected either with Photofrin or with ALA 3-5h before they were killed. The organs were removed directly after and snap-frozen in carbon dioxide ice with isopentane. No further staining or fixation procedures were adopted.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 245-253.
Modelling of inclined and curved surfaces in the reflection scanning acoustic microscope
W. Weise, P. Zinin & S. Boseck Institute for Materials Science and, Structure Research, Physics Department, University of Bremen, 28334 Bremen, Germany
SUMMARY
An expression is derived for the output signal when an inclined plane surface is imaged by the reflection scanning acoustic microscope, which is modelled as a spherical transducer. This expression is applied to model non-planar surfaces. The accuracy of this approach is tested for perfectly reflecting spherical surfaces. The influence of inclination on V(z) curves is considered when Rayleigh waves occur.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 254-261.
Scanning force microscopy on live cultured cells: imaging and force-versus-distance investigations
D. Ricci & M. Grattarola Dipartimento di Ingegneria Biofisica, ed Elettronica, Universita degli Studi di Genova, Via Opera Pia 11a, 16145 Genova, Italy
SUMMARY
Extensive measurements with the scanning force microscope (SFM) on living cells in their native liquid environment are described with the purpose of critically assessing the extent of the interaction between the SFM tip and the (soft) cell materials and the effect of such interaction on topographic information. Images are obtained under various force conditions and systematically correlated with force-versus- distance curves. As a result, detailed indications about tip indentation are given, thickness estimates deduced and identification of submembranous cytoplasmic structures suggested.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 262-275.
In vivo analysis of angiogenesis and revascularization of transplanted pancreatic islets using confocal microscopy
F. A. Merchant, S. J. Aggarwal, K. R. Diller & A. C. Bovik Biomedical Engineering Research Program, ENS 612, University of Texas, Austin 78712-1084, U.S.A.
SUMMARY
A technique to measure angiogenesis and revascularization in pancreatic islets transplanted at the renal subcapsular site in the rat has been developed. In vivo imaging of the microcirculation of transplanted pancreatic islets was conducted using a confocal scanning laser microscope (CSLM) to achieve optical sectioning through the graft in order to perform a computer reconstruction of the three-dimensional neovascular morphology. Individual islets were harvested by enzymatic digestion of excised pancreas from Fischer 344 rats. Isolated islets were cultured for 24h, and approximately 300- 350 islets were transplanted at the renal subcapsular site of the left kidney in an anaesthetized rat. Six to 14 days post- transplantation, the animal was anaesthetized and prepared for in vivo imaging of the microvasculature on a Zeiss LSM-10. Optical contrast of the microvasculature was enhanced by the administration of fluorescein-labelled dextran into the circulating blood. The transplant site was identified and serial sections were obtained through the vascular bed at varying z-intervals. Complementary fluorescence video images were also obtained via a silicon intensifier tube camera mounted on the CSLM. At completion of the imaging procedure, the kidney was returned into the body cavity, the area was sutured and the animal was allowed to recuperate for subsequent examinations. Image processing algorithms, such as grey-level thresholding, median filtering, skeletonization and template matching, were applied to compute the vessel density and diameters and extrapolated to measure 3-D vessel lengths and the tortuosity index of the neovasculature.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 276-280.
Optoelectronic detector probes for scanning near-field optical microscopy
H. U. Danzebrink Physikalisch-Technische Bundesanstalt, Bundesallee 100, D-38116 Braunschweig, Germany
SUMMARY
A brief explanation of the optoelectronic probe concept and a comparison between the implementation of passive waveguide probes and optoelectronic probes in scanning near-field optical microscopy (SNOM) is presented. The first probe realizations using cleaved semiconductor crystals and the work at present in progress using microfabricated Si pyramids are described. These crystals with evaporated metal electrodes forming a slit aperture with subwavelength dimensions work as metal-semiconductor-metal photodetectors. Their optical detection behaviour is investigated by measuring the intensity distribution of a laser focal point. Measurements where the external bias voltage is changed show that it is possible to modify the detection behaviour of the device because of the varying depletion widths. The last part of the article describes a concept where pyramidal probes should function simultaneously as senors for scanning force microscopy (SFM) to measure topography and as optoelectronic probes for scanning near-field optoelectronic microscopy (SNOEM).
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 281-286.
Imaging in the far-red with electronic light microscopy: requirements and limitations
C. Cullander School of Pharmacy S926, University of California, San Francisco, CA 94143-0446, U.S.A.
SUMMARY
The acquisition of simultaneous dual confocal images with red and far-red light has both advantages (e.g. lower autofluorescence) and limitations. An understanding of these requisites is necessary to acquire high-quality images and to avoid the misinterpretation of experimental data. The poor detection of far-red light mandates a high optical transfer efficiency for the system, thus the transmittance of the objective lens and its axial and lateral chromatic aberration in the far-red are important factors for consideration. This technical note is an attempt to 'demystify' the process of filter set design for confocal microscopy by discussing the considerations that went into the construction of a filter set for use with the reagents cyanine 3.18 (Cy3) and cyanine 5.18 (Cy5), and thus to encourage users to look beyond the multipurpose designs available commercially. The 568-nm laser line exciting Cy3 is at its emission maximum, which limits the collectable Cy3 fluorescence. High-transmission optical filters with sharp bandpass cutoffs are thus desirable for maximum light throughput. Light path mirror efficiency rapidly degrades above 700nm, but the loss of this portion of the Cy5 emission spectrum is acceptable since the fluorophore is very bright, and these very long wavelengths are also likely to introduce aberration. While resolution is decreased with far- red light, there is also greater penetration and less scattering, and it is thus possible to obtain high-quality images from deeper within the specimen. Although only one make and model of confocal microscope (the Bio-Rad MRC-600) is considered, similar considerations pertain to the design of filter sets for any confocal microscope that accommodates user-installed filters.
Journal of Microscopy, Vol. 176, Pt. 3, December 1994, pp. 287-299.
Simultaneous confocal recording of multiple fluorescent labels with improved channel separation
K. Carlsson, N. Aslund, K. Mossberg & J. Philip Physics IV, The Royal Institute of Tech, S-100 44 Stockholm, Sweden
SUMMARY
Confocal microscopes are often used to study specimens labelled with fluorophores. A commonly used method for simultaneous recording of the distribution of multiple fluorophores is to divide the fluorescent light emitted by the specimen into different wavelength regions using dichroic and bandpass filters. These different wavelength regions are then distributed to multiple detectors. However, fluorophores often result in considerable cross-talk between channels. A new technique, intensity-modulated multiple-beam scanning (IMS) microfluorimetry, can be used to reduce this cross-talk substantially. The IMS technique is implemented with two laser beams of different wavelengths, intensity-modulated at different frequencies, which illuminate the specimen simultaneously. The two laser wavelengths predominantly excite one fluorophore each. Fluorescent light from the specimen is divided into two wavelength regions (red and green) which are detected by two photomultiplier tubes. The output signals from the photomultiplier tubes are connected to lock-in amplifiers. The effect of using modulated laser beams, in combination with lock-in amplifiers, is strongly to reduce the cross-talk between channels. The performance of the IMS technique using various types of specimen is compared with the results obtained using the conventional multi-detector design.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 1-6.
In vivo determination of fibril orientation in plant cell walls with polarization CSLM
J.-P. Verbelen & D. Stickens Department of Biology, University of Antwerp (UIA), Universiteitsplein 1, B-2610 Wilrijk-Antwerpen, Belgium
SUMMARY
Congo Red fluorescence is used to detect cellulose in the wall of plant cells. The orientation of the cellulose fibrils is determined by using polarized light for excitation. The absorption characteristics of Congo Red make this approach a method of choice for applications with any standard confocal scanning laser microscopy (CSLM). The semi-quantitative character of CSLM observations combined with the non-toxicity of the stain allow a very fast and reliable assessment of cellulose orientation in the wall of living plant cells.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 7-17.
Three-dimensional reconstruction of the human renal glomerulus
K. Preston Jr, B. Joe, R. Siderits & J. Welling Kensal Corporation, 5055 East Broadway (Suite C206), Tucson AZ 85711, U.S.A.
SUMMARY
The capillary bed of human renal glomerulus is one of the more complex capillary structures in the human body. This paper illustrates three-dimensional reconstruction of the capillary bed from serial sections. It shows that, although traditional methods of three-dimensional rendering by computer fail to handle the complexities of the capillary structure, new methods based on filtering using three-dimensional mathematical morphology are capable of revealing previously unseen details. This is done at the expense of eliminating fine structure (small capillaries). An error analysis allows the degree to which fine details are lost to be estimated.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 18-30.
Quantitative water mapping of cryosectioned cells by electron energy loss spectroscopy
S. Q. Sun, S.-L. Shi, J. A. Hunt & R. D. Leapman Health & Human Services, Public Health Service, National Institutes of Health, Building 13 Room 3W13, Bethseda MD 20892, U.S.A.
SUMMARY
A direct technique based on electron energy-loss spectroscopy (EELS) in the scanning transmission electron microscope (STEM) has been developed to map subcellular distributions of water in frozen-hydrated biological cryosections. Previously, methods for water determination have been indirect, in that they have required the cryosections to be dehydrated first. The new approach makes use of spectrum-imaging, where EELS data are collected in parallel at each pixel. Several operations are required to process the spectra including: subtraction of the detector dark current, deconvolution by the detector point-spread function, removal of plural inelastic scattering and correction for the support film. The resulting single scattering distributions are fitted to standard reference spectra at each pixel, and water content can be determined from the fitting coefficients. Although the darkfield or brightfield image from a hydrated cryosection shows minimal structure, the processed EELS image reveals strong contrast due to variation in water content. Reference spectra have been recorded from the major biomolecules (Protein, lipid, carbohydrate, nucleic acid) as well as from vitrified water and crystalline ice. It has been found that quantitative results can be obtained for the majority of subcellular compartments by fitting only water and protein reference spectra, and the accuracy of the method for these compartments has been estimated as plus/minus 3.5%. With the present instrumentation the maximum allowed dose of 2000e/nm2 limits the useful spatial resolution to around 80nm plus/minus 5% precision at a single pixel. By averaging pixel intensities a value of 56.8% with a precision of plus/minus 2.0% has been determined for the water content of liver mitochondria. The water mapping technique may prove useful for applications to cell physiology and pathophysiology.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 31-42.
Hybrid scanning transmission electron/scanning tunnelling microscope system for the preparation and investigation of biomolecules
H. F. Knapp, R. Wyss, R. Haring, C. Henn, R. Guckenberger & A. Engel Maurice E Muller Institut fur, Hochauflosende Elektronenmikroskopie, Universitat Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland
SUMMARY
A hybrid scanning transmission electron/scanning tunnelling microscope vacuum system is introduced, which allows freeze drying and metal coating of biological samples and their simultaneous observation by scanning transmission electron microscopy and scanning tunnelling microscopy (STM). Different metal coatings and STM tips were analysed to obtain the highest possible resolution for such a system. Bovine liver catalase was used as a test sample and the STM results are compared to a molecular scale model.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 43-52.
Backscattered electron imaging of the undersurface of resin-embedded cells by field emission scanning electron microscopy
R. G. Richards & I. Ap Gwynn AO/ASIF Research Institute, Clavadelerstrasse, CH-7270 Davos Platz, Switzerland
SUMMARY
In this study backscattered electron (BSE) imaging was used to display cellular structures stained with heavy metals within an unstained resin by atomic number contrast in successively deeper layers. Balb/c3T3 fibroblasts were cultured on either 13mm discs of plastic Thermanox, commercially pure titanium or steel. The cells were fixed, stained and embedded in resin and the disc removed. The resin block containing the cells was sputter coated and examined in a field-emission scanning electron microscope. The technique allowed for the direct visualization of the cell undersurface and immediately overlying areas of cytoplasm through the surrounding embedding resin, with good resolution and contrast to a significant depth of about 2 micrometre, without the requirement for cutting sections. The fixation protocol was optimized in order to increase heavy metal staining for maximal backscattered electron production. The operation of the microscope was optimized to maximize the number of backscattered electrons produced and to minimize the spot size. BSE images were collected over a wide range of accelerating voltages (keV), from low values to high values, to give 'sections' of information from increasing depths within the sample. At 3- 4keV only structures a very short distance into the material were observed, essentially areas of cell attachment to the removed substrate. At higher accelerating voltages information on cell morphology, including in particular stress fibres and cell nuclei, where heavy metal were intensely bound, became more evident. The technique allowed stepwise 'sectional' information to be acquired. The technique should be useful for studies on cell morphology, cycle and adhesion with greater resolution than can be obtained with any light-microscope- based system.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 53-67.
Tomographic reconstruction of the cross-sectional refractive index distribution in semi-transparent birefringent fibres
T. C. Wedberg & W. C. Wedberg Physics Department, University of Bergen, Allegt 55, N-5007 Bergen, Norway
SUMMARY
Optical diffraction tomography (ODT) is used to reconstruct the complex refractive index distribution in cross-sections of semi-transparent, birefringent fibres. The selected fibres were polymer and animal fibres of either circular or non- circular cross-section with average thicknesses in the range 8-110 micrometre. This choice of samples was made to illustrate the imaging capabilities of ODT, and also to demonstrate some potential applications of the technique. The images representing the reconstructed refractive index distributions have a spatial resolution of about 2 micrometre, and show noticeable image contrast for refractive index variations of about 0.001. The ODT reconstructions compare well with refractive index information provided with the samples, and with scanning electron micrographs of cross- sections of the same fibre samples. From these results it appears that ODT can be used to reconstruct the complex refractive index distribution in cross-sections of semi- transparent birefringent fibres.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 68-76.
Computer simulated high-resolution transmission electron microscopy (HRTEM) in tourmaline
E. A. Ferrow Avdelningen for Mineralogi och Petrologi, Geologiska Institutionen, Lund Universitet, Solvegatan 13, 223 62 Lund, Sweden
SUMMARY
The contrast distributions observed in high-resolution transmission electron microscopy (HRTEM) images of tourmaline depend on the types and magnitudes of the exchange components present and on the degree of atom overlap along the direction of observation. Furthermore, the fractional atomic coordinates in tourmalines are valid only for the specific specimen refined. These properties make the interpretation of experimental HRTEM images of tourmaline using image simulation if not impossible at least extremely difficult. A correct interpretation of experimental HRTEM images of tourmaline is possible provided the structural refinement data on the same crystal are available. Nevertheless, it is possible to interpret the experimental HRTEM images of tourmaline if the composition of the structural model chosen during image simulation approximates the composition of the specimen studied by electron microscopy. A good control of the composition of the specimen studied and an appropriate choice of a structural model for image simulation are therefore as important as properly controlling specimen thickness, specimen tilt, beam tilt and objective lens defocus.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 77-84.
Confocal microscopy in the analysis of the etched nuclear particle tracks in polymers
J. Jakes, P. Gais & H. Schraube Institut fur Strahlenschutz, GSF-Neuherberg, Postfach 1129, D-85758 Oberschleissheim, Germany
SUMMARY
The possibility of the morphometric analysis of etched tracks, induced by protons and alpha particles in the organic polymer allyl diglycol carbonate (CR-39), using the confocal scanning laser microscope (CSLM), was studied. The detectors were investigated in two groups of irradiation experiments, namely: (a) irradiated with mono-energetic neutrons of energy 1.2MeV, (b) exposed to the alpha radiation from 222Rn and its progeny. Both groups were irradiated at normal incidence. Radiation- induced latent tracks were electrochemically etched, and their morphometric parameters were investigated in the reflection mode by using the 488nm spectral line of an argon ion laser. A constant number of up to 200 optical sections in Z-scan mode was taken through each selected etched track at vertical spacings of 0.642 micrometre. Successive reconstructions of Z- sections were used to determine the following parameters: the mean radius of the opening channel, the maximum diameter and the length of the track, and the angle of the track wall to the surface of the sample. The results show that tracks produced by alpha particles differ from those induced by protons. The radius of the opening channel of alpha-particle- induced tracks ranges from 7.9 to 11 micrometre, whereas for protons the same parameter ranges between 2.0 and 3.8 micrometre for a specific electrochemical etch procedure.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 85-89.
A simple method for overcoming some problems when observing thick reflected biological samples with the confocal scanning laser microscope
C. Rumio, M. Morini, J. R. Miani, I. Barajon & P. Castano Institute of Human Anatomy, Via Mangiagalli 31, 20133 Milano, Italy
SUMMARY
A simple device is described, which allows the range of depth of scanning to be reduced when observing thick reflecting biological specimens with a confocal scanning laser microscope (CSLM). Thick histological sections of human skin and rat brain stem were mounted between two coverslip ('sandwich style') and the optical tomography was performed from both sides by turning the 'sandwich' upside-down. The samples were impregnated using standard Golgi-Cox, 'rapid Golgi' or other silver methods. The ability to turn the sandwich upside-down is particularly useful when the reflective structure inspected is deep inside the section, i.e. near the lower surface of the specimen, or when it is opaque to the laser beam of excessively reflective.
Journal of Microscopy, Vol. 177, Pt. 1, January 1995, pp. 90-92.
Thick section preparation using a silicon-rubber-based sealant
H. Cox, C. Walker & C. V. Howard Department of Orthopaedic & Accident Surgery, Royal Liverpool University Hospital, Prescot Street, PO Box 147, Liverpool, L69 3BX
SUMMARY
A method has been developed, using a silicon-rubber-based sealant, which allows 2-3-mm-thick specimens to be maintained in a protected fluid environment for a number of months, without risk of dehydration. Following this, the specimen can be retrieved, stained, embedded and sectioned further. For example, 2-mm-thick sections of fixed unstained bone are easily examined by means of epi-illuminated polarized light and fluorescence microscopies using either conventional or confocal optics. The method could easily be extended to other tissues, for example brain tissue.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
A method to compensate for light attenuation with depth in three-dimensional DNA image cytometry using a confocal scanning laser microscope
A. Liljeborg, M. Czader & A. Porwit Physics IV, The Royal Institute of Tech, S-100 44 Stockholm, Sweden
SUMMARY
A method to compensate for attenuation of detected light with increased depth of the collected optical section, and its application in three-dimensional (3-D) DNA image cytometry is described. The method is based on studying the stack of 2-D histograms that ca be formed from each consecutive pair of sections in a stack of optical serial sections. An attenuation factor is calculated interactively and a new compensated section series is computed. Formalin-fixed paraffin-embedded rat tissue was stained with propidium iodide. Each cell nucleus is extracted by thresholding and its total intensity is calculated. The coefficient of variation (CV) of the total intensity of all cells in each stack is computed. For comparison the CV of the same cells is computed in the uncompensated stacks. This study shows a significantly lower CV for the compensated data, thus contributing to the accuracy of DNA quantification in 3-D DNA image cytometry.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
Microfractography of granitic rocks under confocal scanning laser microscopy
M. Montoto, A. Martinez-Nistal, A. Rodriguez-Rey, N. Fernandez-Merayo & P. Soriano University of Oviedo, Department of Geology, Group of Petrophysics, 33005 Oviedo, Spain
SUMMARY
Scanning laser microscopy, in the confocal mode (CSLM) has been applied to a granitic rock to characterize its fissure space. The technique provides a unique three-dimensional picture of the rock microfractomography. CSLM is unique in observing fine details of the fractographic network (connectivity, tortuosity, etc.), its geometry and its relation to other rock-forming components. The fractographic images with standard fluorescence microscopy are compared with those obtained with CSLM. The examples presented emphasize the advantages of CSLM: three- dimensional visualization of the microfractographic network, crack connectivity, automatic evaluation of direction and slope of fissures. These studies are related to the migration of radionuclides in the geosphere. The relations between potentially water-conducting open fissures and the rock- forming minerals provide a means of modelling the 'radionuclide retardation mechanism', a security factor in their definitive storage in rock masses.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
PHOEBE, a prototype scanning laser-feedback microscope for imaging biological cells in aqueous media
T. L. Wong, S. L. Sabato & A. Bearden Division of Neurobiology, Department of Molecular & Cell Biology, 229 Stanley Hall, University of California at Berkeley, Berkeley CA 94720-3206, U.S.A.
SUMMARY
Based on the principle of laser-feedback interferometry (LFI), a laser-feedback microscope (LFM) has been constructed, capable of providing an axial (z) resolution of a target surface topography of approximately 1nm and a lateral (x,y) resolution of approximately 200nm when used with a high- numerical-aperture oil-immersion microscope objective. LFI is a form of interferometry in which a laser's intensity is modulated by light re-entering the illuminating laser. Interfering with the light circulating in the laser resonant cavity, this back-reflected light gives information about an object's position and reflectivity. Using a 1-mW He-Ne (wavelength=632.8nm) laser, this microscope (PHOEBE) is capable is obtaining 256x256-pixel images over fields from 10x10 micrometre to 120x120 micrometre in approximately 30s. An electrochemical feedback circuit holds the optical pathlength between the laser output mirror and a point on the scanned object constant; this allows two types of images (surface topography and surface reflectivity) to be obtained simultaneously. For biological cells, imaging can be accomplished using back-reflected light originating from small refractive-index changes (} 0.02) at cell membrane/water interfaces; alternatively, the optical pathlength through the cell interior can be measured point-by-point by growing or placing a cell suspension on a higher-reflecting substrate (glass or silicon wafer). Advantages of the laser-feedback microscope in comparison to other confocal optical microscopes include: simplicity of the single-axis interferometric design; the confocal property of the laser-feedback microscope (a virtual pinhole), which is achieved by the requirement that only light that re-enters the laser meeting the stringent frequency, spatial (TEM00), and coherence requirements of the laser cavity resonator mode modulate the laser frequency; and the improved axial resolution, which is based on interferometric measurement of optical amplitude and phase rather than by use of a pinhole as in other types of confocal microscopes.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
Imaging periodic surface relief structures
J. T. Sheridan & T. O. Korner TP 680, Institute for System Engineering and, Informatics Science R&D, Joint Research Centre JRS/CCR, I-21020 Ispra (VA), Italy
SUMMARY
Because of shadowing, multiple scatter and polarization effects, the interpretation of images of grating with fine periods, isolated deep structures, and multiple scattering volume objects is seriously complicated. In this paper a review of methods used to model such effects is presented. Periodic surface relief gratings are of particular current importance because of the possibility of producing calibration samples using them. Several examples which illustrate electromagnetic volume effects are examined. General trends which help in validating the use of Fourier-transform-based scalar transmittance theory are then indicated. The angular spectrum approach, which can be used , together with a scatter function generated using the rigorous electromagnetic theory, to calculate coherent, partially coherent and confocal images of volume objects, is also discussed.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
Application of confocal laser microscopy and three-dimensional Voronoi diagrams for volume and surface area estimates of interphase chromosomes
R. Eils, E. Bertin, K. Saracoglu, B. Rinke, E. Schrock, Y. Usson, M. Robert-Nicoud, E. H. K. Stelzer, J.-M. Chassery, T. Cremer & C. Cremer Institut fur Angewandte Physik, Albert-Uberle Strasse 3-5, D-69120 Heidelberg, Germany
SUMMARY
This study demonstrates the use of Voronoi tessellation procedures to obtain quantitative morphological data for chromosome territories in the cell nucleus. As a model system, chromosomes 7 and X were visualized in human female amniotic fluid cell nuclei by chromosomal in situ suppression hybridization with chromosome-specific composite probes. Light optical serial sections of 18 nuclei were obtained with a confocal scanning laser fluorescence microscope. A three- dimensional (3-D) tessellation of the image volumes defined by the stack of serial sections was then performed. For this purpose a Voronoi diagram, which consists of convex polyhedra structured in a graph environment, was built for each nucleus. The chromosome territories were then described by three morphological parameters, i.e. volume, surface area and a roundness factor (shape factor). The complete evaluation of a nucleus, including the calculation of the Voronoi diagram, 3-D visualization of extracted territories using computer graphic methods and parameterization was carried out on a Silicon Graphics workstation and was generally completed within 5 min. The geometric information obtained by this procedure revealed that both X- and 7-chromosome territories were similar in volume. Roundness factors indicated a pronounced variability in interphase shape for both pairs of chromosomes. Surface estimates showed a significant difference between the two X- territories but not between chromosome 7-territories.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
H. Heinzelmann, B. Hecht, L. Novotny & D. W. Pohl Institut fur Physik, Universitat Basel, Klingelbergstrasse 82, 4056 Basel, Switzerland
SUMMARY
Near-field optics (NFO) opens the door to light microscopy techniques with resolutions well beyond the diffraction limit. The richness of optical investigations is now applicable on a near-molecular level. Among the novel scanning near-field optical microscopy (SNOM) schemes, the most prominent representatives are aperture SNOM and scanning tunnelling optical microscopy (STOM or PSTM). New experimental and theoretical work has to be performed to study the phenomena specific to NFO. One such example is the angular dependence of light emission in aperture SNOM. The detection of radiation at angles greater than the critical angle of total internal reflection alpha=arcsin(1/n), where n is the sample refractive index, can represent a microscopy scheme that combines the respective advantages of both aperture SNOM and STOM. Recent experiments have demonstrated the expected exponential dependence of light intensity on gap width (for fixed emission angle). The decay length as a function of alpha is in agreement with the Fresnel description of the evanescent field when total reflection occurs at an interface. These investigations were additionally motivated by calculations based on the multiple multipole method.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
Automated correction of linear deformation due to sectioning in serial micrographs
T. Jansson, T. Gustavsson, M. Rydmark, C.-H. Berthold, R. Pascher & T. Skoglund Department of Applied Electronics, Chalmers University of Technology, S-412 96 Goteborg, Sweden
SUMMARY
This paper describes an objective and automatic method for detection and correction of sectioning deformations in digitized micrographs, as well as an evaluation of the method applied to light and electron microscopic images of semi-thin and ultra-thin serial sections from brain cortex. The detection is based on matching of image subregions and the deformation model is bi-linear, i.e. two first-order polynomials are used for modelling compression/expansion in perpendicular directions. The procedure is applicable to prealigned serial two-dimensional sections and is primarily aimed at three-dimensional reconstruction of tissue samples consisting of a large number of cells with random distribution and morphology.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
Comparison of three-dimensional imaging properties between two-photon and single-photon confocal fluorescence microscopy
Min Gu & C. J. R. Sheppard Physical Optics Department, School of Physics, The University of Sydney, NSW 2006, Australia
SUMMARY
The imaging performance in single photon (1-p) and two-photon (2-p) fluorescence microscopy is described. Both confocal and conventional systems are compared in terms of the three- dimensional (3-D) point spread function and the 3-D optical transfer function. Images of fluorescent sharp edges and layers are modelled, giving resolution in transverse and axial directions. A comparison of the imaging properties is also given for a 4Pi confocal system. Confocal 2-p 4Pi fluorescence microscopy gives the best axial resolution in the sense that its 3-D optical transfer function has the strongest response along the axial direction.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
Double pulse fluorescence lifetime imaging in confocal microscopy
M. Muller, R. I. Ghauharali, K. Visscher, T. D. Visser & G. J. Brakenhoff Department of Molecular Cytology, University of Amsterdam, Plantage Muidergracht 14, 1018 TV Amsterdam, The Netherlands
SUMMARY
A theoretical analysis of a new technique for fluorescence lifetime measurement, relying on (near steady state) excitation with short optical pulses, is presented. Application of the technique to confocal microscopy enables local determination of the fluorescence lifetime, which is a parameter sensitive to the local environment of fluorescent probe molecules in biological samples. The novel technique provides good time resolution, since it relies on the laser pulse duration, rather than on electronic gating techniques, and permits, in combination with bilateral confocal microscopy and the use of a (cooled) CCD, sensitive signal detection over a large dynamic range. The principle of the technique is discussed within a theoretical framework. The sensitivity of the technique is analysed, taking into account: photodegradation, the effect of the laser repetition rate and the effect of non-steady-state excitation. The features of the technique are compared to more conventional methods for fluorescence lifetime imaging.
Journal of Microscopy, Vol. 177, Pt. 2, February 1995, pp. .
Near field imaging: some attempts to define an apparatus function
D. Courjon Laboratoire d'Optique PM Duffieux, Associe au CNRS URA-214, UFR des Sciences et des Techniques, 25030 Besancon Cedex, France
SUMMARY
Near-field microscopy is a promising new tool capable of imaging details smaller than the wavelength. The mechanism of imaging is analysed and an overview of the apparatus functions which could be used to define an image quality criterion is given.
Apologies for sending such a long file at once. Please note that summaries will be posted less frequently as Gillian Wilson is now on maternity leave.
Gillian Wilson Executive Editor The Royal Microscopical Society Journal of Microscopy & 37/38 St Clements Proceedings of the RMS Oxford OX4 1AJ UNITED KINGDOM rms-at-uk.ac.ox.vax OR rms-at-vax.ox.ac.uk Telephone +44 (0)865 248768 Fax +44 (0)865 791237
I apologize for not being complete regarding the URL I gave on the SCIL-Image software. Here is a more precise specification of the path towards the information.
URL: http://www.tn.tudelft.nl
select : Pattern Recognition Group - Software Developments - SCIL-Image
Hope this saves you time.
-- Kees van der Wulp
******************************* ATTENTION *************************** * * * Change in INTERNET address ! * * * * From : vanderwulp-at-mbl.tno.nl * * To : vanderwulp-at-voeding.tno.nl * * * *********************************************************************
TNO - Nutrition & Food Research INTERNET : vanderwulp-at-voeding.tno.nl Division : Toxicology VOICE : +31 15 843101 Department : Genetic Toxicology FAX : +31 15 843989 PO-Box 5815 General TNO Info : http://www.tno.nl 2280 HV RIJSWIJK (NL) THE NETHERLANDS
Groeten, Gert van Antwerpen. TNO Institute of Applied Physics. P.O.Box 155, 2600 AD Delft, The Netherlands. Phone: +31 15-692226, Fax: +31 15-692111 Email: antwerp-at-tpd.tno.nl
-- Kees van der Wulp
******************************* ATTENTION *************************** * * * Change in INTERNET address ! * * * * From : vanderwulp-at-mbl.tno.nl * * To : vanderwulp-at-voeding.tno.nl * * * *********************************************************************
TNO - Nutrition & Food Research INTERNET : vanderwulp-at-voeding.tno.nl Division : Toxicology VOICE : +31 15 843101 Department : Genetic Toxicology FAX : +31 15 843989 PO-Box 5815 General TNO Info : http://www.tno.nl 2280 HV RIJSWIJK (NL) THE NETHERLANDS
1,4-Diazobicyclo(2,2,2)octane (Dabco) delays fading of immunofluorescence preparations Langanger, Gabriele; De Mey, Jan; Adam, Hans Mikroskopie 1983; 40 (7-8): 237-41 Language: German
Fading of immunofluorescence during microscopy. A study of the phenomenon and its remedy Johnson G D; Davidson R S; Mcnamee K C; Russell G; Goodwin D; Holborow E J J Immunol Methods 55 (2). 1982. 231-242
Regards, Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
I encountered the identical problem of dark caramel colored epon mix after addition of DMP-30, Substitution of the standard DDSA component with a new bottle of "Specially Distilled DDSA" corrected the problem and produced the golden colored resin mix. I was also told (see Marge Hukee's reply) the problem was a function of the pH of the resins
---------------------------------------------------------- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego CA 92182-0057 phone: (619) 594-4523 fax: (619) 594-5676 email to sbarlow-at-sunstroke.sdsu.edu
Because I still miss the original message and only received the additional message I posted (Re : 2D software for Unix), I repost the original correc- ted message to the microscopy list. I apologize for this 2nd message if you DID receive the first one somehow.
Dear Norm,
Regarding your question,
} } I ask for comments, recommendations, advice, for 2D image analysis software } for a UNIX platform (SGI Indy R4000). } } If there is interest I'll post a summary. Thanks. }
I can give you 2 URL's, leading to the same SCIL page, that provide you with a brief description of SCIL-Image (version 1.3), as well as an e-mail address of Mr. G. van Antwerpen whom I gave your address. He will take care of providing you with more information on SCIL-Image.
I am using SCIL-Image on my Personal IRIS 4D/35 for a couple of years now. It is an extensive 2D multi-layered image processing package (} 200 ip commands). It can be used on various platforms (Sun, HP, SGI, IBM, PC's, Mac's) and can be run from a mouse driven menu(1), a commandline interpreter (2) and from compiled routines (3) which makes it run very fast. It is a combination of a Standard C Interpreter Language (part 1) and an extensive Image processing library (part 2). It incorporates a complete program flow control mechanism and enables you to create and mix C- statements (and complete programs) with image processing commands. You can also easily extend the package with your own library of user created (compiled or C-source) routines and incorporate them into the mouse driven menu. I run it from my X-Terminal (Tektronix XP 27 and XP 356, colour terminals) in interactive as well as batch mode. I heard from mr. van Antwerpen that you can evaluate the full package for a period of 3 months at a rate of about US$ 250.= However for these kind of things please contact him, for I am only a user of the package.
URL: http://www.tn.tudelft.nl
select : Pattern Recognition Group - Software Developments - Scil-Image
or URL: http://galaxy.ph.tn.tudelft.nl:2000/Software/scil.html
e-mail address G. van Antwerpen : antwerp-at-tpd.tno.nl
All people interested in more info can get it from him.
Good luck,
-- Kees van der Wulp
TNO - Nutrition & Food Research INTERNET : vanderwulp-at-voeding.tno.nl Division : Toxicology VOICE : +31 15 843101 Department : Genetic Toxicology FAX : +31 15 843989 PO-Box 5815 General TNO Info : http://www.tno.nl 2280 HV RIJSWIJK (NL) THE NETHERLANDS
We have developed a new method for fast data transfer from a JEOL microscope connected to a LINK eXL computer system to DOS based personal computers.
In our institute we are using a JEOL JSM 6400 SEM equipped with an energy dispersive detector driven by a LINK eXL computer running the GENIE operating system.
In former days we experienced great problems transferring images captured from the screen and stored in tiff format to DOS based computers, since we neither posses any GENIE image processing software nor do we have network connectivity on the Link system.
That's why we have developed a unique hardware/software solution which allows users to transfer tiff files at the rate of approx. 1 sec. per file (262 KB) from the Link computer to the DOS computer.
The software is capable of viewing the contents of GENIE formatted floppy disks, hard disks, MO's and changeable devices (e.g. Syquest drives) and copying selected images and spectra to the target computer.
Anyone interested may contact us for further details at:
I publish a monthly newsletter, Analytical Consumer, that reports on customer satisfaction with manufacturers of analytical equipment. We are currently doing a survey of TEM users, similar to an SEM study we published last July, in conjunction with Microscope News & Technology (who is also collaborating on this study). I am looking for labs with TEMs who would be willing to talk about their equipment.
I'm looking for:
What kind of TEM (manufacturer and model) do you use? How old is it (roughly)? Why did you buy from that particular manufacturer? Have you been satisfied with the equipment? Are you satisfied with the service? Do you get service from the equipment mfr, do it yourself, or have an outside service supplier? Do you have a service contract? What kind of samples do you typically use the TEM for? What kind of lab are you in? R&D, research, instrument facility, service lab, or ...? What kind of company is the lab?
If you answer these questions and want a copy of the survey, please include your address. If you would like to talk about it in person, give me a call at (508) 369-9079, or send me your phone number and I'll call you.
Thanks!
Jo Rita Jordan Editor and Publisher Analytical Consumer
jjordan-at-world.std.com or 76150.2171-at-compuserve.com
MICROSCOPY CONFERENCE (In conjunction with the 18th National Conference of the New Zealand Society for Electron Microscopy)
Dunedin, New Zealand 4th - 8th September 1995.
A Microscopy Conference will be held in Dunedin, 4th to 8th September, 1995. The conference will be held in conjunction with the 18th National Conference for the New Zealand Society for Electron Microscopy (NZSEM) however will cover all aspects of Microscopy with emphasis on the techniques of both Light Microscopy and Electron Microscopy.
The venue for the conference will be the Otago Medical School.
Workshop sessions will run on Monday and Tuesday (Sept 4th - 5th); the conference proper will run from Wednesday until midday Friday (Sept 6th - 8th).
Guest speakers include:
Professor John M Robinson, Ohio State University, U.S.A. Professor Robinson has published widely on the practical aspects of enzyme cytochemistry (in particular the recently developed cerium-based techniques) and immunocytochemistry. His application of these techniques utilises many forms of microscopy including conventional light microscopy, confocal microscopy, transmission electron microscopy and scanning electron microscopy.
Professor Anthony S-Y Leong, University of Adelaide, Australia. Professor Leong is well known for his work with microwave techniques, both at the light microscope and the electron microscope level. His application of microwave technology includes fixation and processing for L.M. and T.E.M., and the use of microwaves in immunocytochemistry.
Dr Brian Brooker, Institue of Food Research, England. Dr Brooker is a food scientist particularly interested in the structure and behaviour of oil-in-water emulsions and the influence of emulsifiers on their functions. He applies many microscopy techniques to study these difficult samples including conventional light microscopy, confocal microscopy, freeze fracture, X-ray microanalysis, cryofixation and electron microscopy.
Dr Brendon Griffin, Centre for Microscopy and Microanalysis, Perth, Australia. Dr Griffin's field of interest are the microscopy of rare minerals. He is experienced in microprobe analysis particularly EDS, environmental SEM and field emission SEM. He has a particular interest in EM education. Dr Griffin is currently president of the Australian Society for Electron Microscopy.
Trades displays will be a feature of the Conference. We are also planning a techniques forum with the invited guests forming the panel.
If you are interested in receiving more information about this conference you are invited to contact the organising committee at the address below;
Allan Mitchell Organising Committee, 1995 Microscopy Conference C/- Department of Anatomy and Structural Biology Otago Medical School P.O. Box 913 Tel; National 03 479 7301 International 64 3 479 7301 Dunedin Fax; National 03 479 7254 International 64 3 479 7254 New Zealand. email address; allan.mitchell-at-stonebow.otago.ac.nz
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
I publish a monthly newsletter, Analytical Consumer, that reports on customer satisfaction with manufacturers of analytical equipment. We are currently doing a survey of TEM users, similar to an SEM study we published last July, in conjunction with Microscope News & Technology (who is also collaborating on this study). I am looking for labs with TEMs who would be willing to talk about their equipment.
I'm looking for:
What kind of TEM (manufacturer and model) do you use? How old is it (roughly)? Why did you buy from that particular manufacturer? Have you been satisfied with the equipment? Are you satisfied with the service? Do you get service from the equipment mfr, do it yourself, or have an outside service supplier? Do you have a service contract? What kind of samples do you typically use the TEM for? What kind of lab are you in? R&D, research, instrument facility, service lab, or ...? What kind of company is the lab?
If you answer these questions and want a copy of the survey, please include your address. If you would like to talk about it in person, give me a call at (508) 369-9079, or send me your phone number and I'll call you.
Thanks!
Jo Rita Jordan Editor and Publisher Analytical Consumer
jjordan-at-world.std.com or 76150.2171-at-compuserve.com
I publish a monthly newsletter, Analytical Consumer, that reports on customer satisfaction with manufacturers of analytical equipment. We are currently doing a survey of TEM users, similar to an SEM study we published last July, in conjunction with Microscope News & Technology (who is also collaborating on this study). I am looking for labs with TEMs who would be willing to talk about their equipment.
I'm looking for:
What kind of TEM (manufacturer and model) do you use? How old is it (roughly)? Why did you buy from that particular manufacturer? Have you been satisfied with the equipment? Are you satisfied with the service? Do you get service from the equipment mfr, do it yourself, or have an outside service supplier? Do you have a service contract? What kind of samples do you typically use the TEM for? What kind of lab are you in? R&D, research, instrument facility, service lab, or ...? What kind of company is the lab?
If you answer these questions and want a copy of the survey, please include your address. If you would like to talk about it in person, give me a call at (508) 369-9079, or send me your phone number and I'll call you.
Thanks!
Jo Rita Jordan Editor and Publisher Analytical Consumer
jjordan-at-world.std.com or 76150.2171-at-compuserve.com
Microscopy List {microscopy-at-aaem.amc.anl.gov} , Confocal Microscopy List {confocal-at-ubvm.bitnet} Message-Id: {Pine.3.05.1.9502011127.A7624-9100000-at-pauline.sdsc.edu} Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII
I am in need of access to a confocal scope in the San Diego area. Would anyone know of a lab that has one and who to contact? Thanks in advance for your help.
Marc
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
Jan Coetzee asks about electronic vs. "dog-eared diary" for schedules. Dear Jan, We use a large desk calendar ourselves. It seems easier, since there are frequent changes, and everyone can check it instantly. I'm sure that a calendar page on a computer might do as well, but when a user calls, we are not always booted up, so it would take us more time than it does with paper and pencil. If your scopes are computer-controlled, it might save some time and/or effort to have schedules on the same computer--e.g. the computer could dial a user you need to contact re schedule changes--but we have not thought this to be worthwhile for us. Sometimes low-tech works very well! Yours, Bill Tivol
Jan Coetzee inquired about mfgrs of pltrs. Dear Jan, Melcor makes them. Their address, phone & fax are 1040 Spruce Street Trenton NJ 08648-4587 United States of America (our USA) (609) 393-4178 (phone) (609) 393-9461 (fax) One potential problem is that Peltiers have a limited delta-T, and can get very expensive. I asked Melcor about setting up a cold stage for epitax- ial deposition in a vacuum, and there was nothing which would get cold enough. Furthermore, if you wish to thermostat the device, you need to use a cooler which has a 100% duty cycle and output current proportional to the difference between target and actual temperatures. We found this out when we installed Peltiers in our darkroom as heaters/coolers to maintain developer temperature. The intermittant current put out by the usual commercially available devices blew the Peltiers out! Otherwise, Peltiers are marvelous devices. Good luck. Yours, Bill Tivol
I've received this request for help on locating micrographs of bacteria. I could not help this individual. So is there anyone out there that can?
Cheers....Nestor Z. Your Friendly Neighborhood SysOp =====================================
P.S.
I'm working on concept of adding an Image Library to the Software Library. When it's ready for contributions I'll post a message to the Listserver....
====================================
I need an electron microscope picture of Methanosarcina barkeri and Methanobacterium wolfei. I called the University of California at Berkeley and they gave me your address. They said that they don't keep a file of their past work and would charge $300 per picture to do the work again. I'm fairly certain that the pictures already exist somewhere. I believe that NASA may have some of them but I don't know who to ask there . Thank you for any help in getting any pictures of the two bacteria.
The following files have been delivered to you - please use the eXtract command in the browser to work with them:
MARSALL.ASC (format: Text) ---------------------------------------------------------------------- Thomas K. Wilson wilsontk-at-MUohio.edu Dept. of Botany Miami University ! Miami was a University when Oxford OH 45056 ! Florida belonged to Spain ! USA 513.529.4210 office 513.529.4243 fax
-------------- Enclosure number 1 ---------------- The following is posted as a favor to Marshall University, and to any interested applicants on the MSA-BBS. Please distribute this message to any and all interested people you may know.
I am not involved with this Supervisor search in any way whatsoever (Other than having promised to post this ASAP). Please contact Dr. W.B. Rhoten directly at the address below (his E-mail address is Rhoten-at-musom01.mu.wvnet.edu)
POSSITION OPEN
SUPERVISOR OF ELECTRON MICRSCOPY FACILITY
To supervise and perform day-to-day operations of the Electron Micrscopy Facility and assist students, research assistants, and investigators. Participate in graduate level EM Course.
Bachelor's degree including courses in biological and physical sciences and at least 3 years experience in EM, or a Masters degree and at least 3 years of relevant experience, or a Ph.D. degree. Qualifications include comprehensive knowledge of EM and ability to enhance educational and research activities. Experience in image processing and computers desired.
Qualified applicants should send a cover letter, resume, and list of at least 2 references to:
Dr. W. B. Rhoten Dept. of Anatomy, Cell & Neurobiology School of Medicine, Marshall University Huntington, WV 25704-9388
For full consideration submit application by March 1, 1995.
MARSHALL UNIVERSITY IS AN AFFERMATIVE ACTION/EQUAL OPPORTUNITY EMPLOYER
One of EM Unit users is studying developing Red deer. The samples we are examining are skull and pedicle (developing antler) taken from the deer at set time intervals over atwo year period. The samples are processed routinely, ie glutaraldehyde fixed,decalcified, OsO4 post fixed, uranyl acetate bloc stain, dehydrated and embedded in Agar 100 epoxy resin.
Our problem is that it now seems to be that the glycogen content is of some significance to the investigation. Unfortunately the above process is not ideal for showing the glycogen. Some recently processed samples using potassium ferrocyanide with the osmium are brilliant however we would like to enhance the glycogen in the previous blocks. Does anyone out there know a method to enhance "unstainable" glycogen in ultrathin sections? Thanks in anticipation.
Allan Mitchell South Campus EM Unit allan.mitchell-at-stonebow.otago.ac.nz
} I am looking for a way to convert my PostScript files into "regular" image } files. Does someone know of such siftwares on either Mac or Unix platforms? } Any information would be appreciated. } Do you mean postscript or encapsulated postscript? If postscript then one of the postscript interpreters available should do it. I use one an a PC called GoScript that outputs to TIFF files as well as printers. I am not sure, but you should take a look at the GNU Ghostscript interpreter that runs on virtually anything - certainly on unix systems. If you mean encapsulated postscript (EPS) then just import the files into a Mac word processor such as WordPerfect on Microsoft Word and then copy and paste to whereever you want to. +------------------------------------+ | Dr Douglas Arrell | | Mechanical Performance and Joining | | Institute for Advanced Materials | | 1755 ZG Petten | | Netherlands | | {ARRELL-at-JRC.NL} | | Tel. (+31) 2246 5287 | | Fax (+31) 2246 1917 | +------------------------------------+
Group - Responding to an inquiry from Nestor, Custom Medical Stock Photo is a company which purchases micrographs and then sells them to publications, etc. They have a considerable inventory - in microscopy, and a number "with" bacteria. Many readers may be interested in contacting them and explore the sale of their micrographs. I have, of course, no financial interest in the company.
Custom Medical Storck Photo, Inc. 3819 North Southport Ave Chicago, IL 60613 Tel.: 312-248-3200 Fax: 312-248-7427
Message-Id: {1995Feb03.100340.2874650620-at-dmcmail.ucsf.edu} To: Microscopy-at-aaem.amc.anl.gov (M)
Subject: Time: 9:45 AM OFFICE MEMO Uranyl Glass/FM Stds. Date: 2/3/95 Dear Microscopists, Does anyone have any uranyl glass, or know where it might be obtained? I have been told that it is no longer manufactured commercially. It might be an excellent "generic" fluorescence microscopy control. Are there any commercially available, pre-mounted fluorescence standards besides "MultiSpeck" from Molecular Probes? They are very convenient, but they are not ideal for our applications as DAPI, fluorescein, and Texas Red specific controls. Unfortunately, Flow Cytometry Standards Co. no longer makes pre-mounted standards. I have been managing the UCSF core flow and image cytometry facility ("Lab for Cell Analysis") for 2 years, but I had no real QC for our 2 occasionally used fluorescence microscopes. Now I need to establish QC protocols for 6 additional multi-user, computerized fluorescence (one scanning confocal) microscopes in the "National Molecular Cytogenetics Resource." I was surprised that so few standards (and journal references) seem to be available. Any suggestions or comments would be greatly appreciated. Thank you very much. Kris Kavanau; kavanau-at-dmc.ucsf.edu
**************************************************************************** John A. Kloetzel, Ph.D. {kloetzel-at-umbc.edu} Department of Biological Sciences University of Maryland Baltimore County (UMBC) Catonsville, MD 21228-5329 USA Phone: (410) 455-2247 or -3913 (Lab) FAX: (410) 455-3875 ****************************************************************************
A few months ago a thread ran on fixation and embedding pepper contaminant artefact in biological ultrathin sections. A colleague of mine read the Pepper Summary I compiled and sent me the following fixation protocols that he has used successfully without pepper. Notice the first one in which glutaraldehyde, phosphate buffer AND OSMIUM are all mixed TOGETHER!
If anyone out there wants a copy of my Pepper Summary, contact me off-line at my e-mail and I will zip a copy out to you.
"1. For fixing cilia in mammalian trachea, I have used an "instant fixation" method using a combination of osmium, phosphate buffer, and glutaraldehyde - in the cold - for many years and have never seen the pepper described in the Pepper Summary you sent to me. Right - all that stuff in the same buffer dumped on the tissue. Works great, but may extract a bit of actin.
"The method I used was one described in a paper by Omnoto and Kung in the J. Cell Biol 87:33-46. I think it uses 50 mM NaPhosphate, pH 7.2, 2% OsO4, 2% glutaraldehyde. Add the Osmium just before you add the tissue and fix on ice for 10 min. If you want, you can remove the black fix after 10 min and add another slug of fix for another 10 min but that is optional.
"Omoto and Kung used it to fix Paramecium and their cilia. I have used it to fix mammalian trachea (with their cilia). The advantage is that it seems to "freeze" cilia in position, as opposed to glutaraldehyde, in which cells actualy swim for a dab before either being fixed or dying (we fix cells, we don't kill them, do we?). The osmium does not penetrate for more than a few cell layers but, with epithelial tissue or single cells, it does not make much difference.
"I have never tried that fix method on Chlamydomonas or Tetrahymena. Over the last year I have done a lot of embedding of Chlamy and have never seen pepper. For those beasts, I find that cacodylate gives the best preservation of the cytoplasm, although others find that phosphate buffer works fine too. I do fix with glutaraldehyde in culture medium (pretty much phosphate buffer), overnight in glut in cacodylate, rinse a few times in cacodylate, then into 0.5-1% OsO4 in cacodylate for 30-60 min on ice, rinse with a few changes of water and into uranyl acetate for a few hours prior to dehydrating in acetone and embedding in epon.
"I've also tried another method in which you fix with glut in phosphate buffer, rinse, then incubate overnight in uranyl acetate (in water) at 70 degrees C. It works on Chlamy and avoids osmium. It is supposed to eliminate the need for poststaining of sections, but I did not find this to be the case. I believe that I once read that the reason for staining sections is to put a dab of stain on structures at the last surface of the plastic that the beam sees before blasting through the objective lens. I don't know, but, for Chlamy, I usually use the more old fashioned method that I gave you above. Pepper does not seem to be one of my problems." ------------------------------------------------------------------------ Contributed to MICROSCOPY by:
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Dear Lucille, I just got a number from Kodak for technical questions, but they could probably direct you to a distributer. It is (800) 242-2424 x19. We usually use a local vendor, National Graphics, but I have not noticed a great range of prices with other vendors. Good luck. Yours, Bill Tivol
I am not aware that "cheap" is an a.k.a. for "reliable". Please be kind to the English language.
On Fri, 3 Feb 1995, Lucille A. Giannuzzi wrote:
} Can anyone recommend a reliable (a.k.a. cheap) U.S. vendor for TEM film? } } Thanks in advance, } Lucille Giannuzzi } } } ************************************************************************* } Lucille A. Giannuzzi, Ph.D. } } Dept. of Mechanical and Aerospace Eng. phone (407) 823-5770 } University of Central Florida fax (407) 823-0208 } 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu } Orlando, FL 32816-2450 USA } ************************************************************************* } } } }
DDoes anyone know about work done on immunolabelling of wood tissue of fruit trees. I'm interested in any kind of information I can get on fixation, embedding and the labelling procedure. I plan to be using ProteinA-colloidal gold to tag the antibody.I'll be using a confocal microscope for this study.If anyone knows any work done in this area could you please send me the references. Thank you in advance. my email address: komminen-at-student.msu.edu
Dear Fellow Microscopists, We have recently acquired a scanning electron microscope with a four crystal Wavelength Dispersive Spectrometer but the associated computer system is rather old and probably not worth re-activating. I know that there is software for the Macintosh, such as "D.T.S.A." and "Flame", for ENERGY Dispersive Spectroscopy but my question is "Is there any (Mac) software out there which can handle W(AVELENGTH)DS spectral acquisition and processing?". Any leads would be very much appreciated. Thanks.
Rick Hall Materials Science Program University of Delaware
Reply to: RE} Uranyl Glass/FM Stds. Hi Kris, Uranium glass slides can be purchased from:
Newport Industrial Glass, Inc. 1631 Monrovia Ave. Costa Mesa, CA 92627 Tel: 714-642-9980 Fax: 714-645-6800 Contact person: Bill Larsen (you can tell him I sent you). Sold as a 6.5x6.5" sheet (you can specify 1 mm height), so you may want to form a "consortium" to have Newport pre-cut a sheet to slide size (nominal extra cost, but your lab only needs one or two slides). If there is a lot of interest, my company may start selling single slides.
As for references and the Shading Correction equation: please see my article in the 11/94 issue of Journal of NIH Research 6(11): 80 (usual Internet disclaimer: yes, that is an ad from my company on the facing page). Also look at Jericevic et al (1989) Methods in Cell Biology 30:47-83.
MutliSpeck beads: The Molecular Probes pre-mounted slide kits should be ideal for DAPI and Fluorescein. I believe they were optimized for Rhodamine, but should still work ok for Texas Red. If your problem is with mounting, Mol. Probes now sells the beads in solution, so you can 'sprinkle' some on your specimens. If you have a different problem with the current MultiSpeck's, Mol. Probes may be able to work something out for you.
Sorry, but I usually buy my reference material from Mol. Probes and don't keep close track of other slide manufacturers.
Sincerely,
Dr. George McNamara Universal Imaging Corporation George_M-at-Image1.com --------------------------------------
Subject: Time: 9:45 AM OFFICE MEMO Uranyl Glass/FM Stds. Date: 2/3/95 Dear Microscopists, Does anyone have any uranyl glass, or know where it might be obtained? I have been told that it is no longer manufactured commercially. It might be an excellent "generic" fluorescence microscopy control. Are there any commercially available, pre-mounted fluorescence standards besides "MultiSpeck" from Molecular Probes? They are very convenient, but they are not ideal for our applications as DAPI, fluorescein, and Texas Red specific controls. Unfortunately, Flow Cytometry Standards Co. no longer makes pre-mounted standards. I have been managing the UCSF core flow and image cytometry facility ("Lab for Cell Analysis") for 2 years, but I had no real QC for our 2 occasionally used fluorescence microscopes. Now I need to establish QC protocols for 6 additional multi-user, computerized fluorescence (one scanning confocal) microscopes in the "National Molecular Cytogenetics Resource." I was surprised that so few standards (and journal references) seem to be available. Any suggestions or comments would be greatly appreciated. Thank you very much. Kris Kavanau; kavanau-at-dmc.ucsf.edu
The organizers of the 28-31 March 1995 Scanning 95 meeting want about 25 student volunteers to run the slide projectors for the sessions in exchange for full meeting registration ($275). Please contact: Mary K. Sullivan FAMS, Inc P.O. Box 832 Mahwah, New Jersy 07430,0832
or leave me a message. Debe Holmberg e-mail {dlholmberg-at-peseta.ucdavis.edu} Lab 916-752-9021 FAX 916-752-4604
I am going to be working on an TEM project that will be under "GLP" . GLP are guidlines for doing certain experiments for the FDA (I think the equivalent in Europe is ISO 9000). I was wondering if anyone out there is doing TEM under these guidlines? And if so what are they using for magnification calibration. I do not believe that there are any vendors who can supply a certified magnification standard for TEM, which, I think is required for GLP.
Thanks, David Leaffer Syntex Research david.leaffer-at-syntex.com
The organizers of the 28-31 March 1995 Scanning 95 meeting want about 25 student volunteers to run the slide projectors for the sessions in exchange for full meeting registration ($275). Please contact: Mrs. Mary K. Sullivan FAMS. Inc. P.O. Box 832 Mahwah, NJ 07430-0832
or leave me a message. Debe Holmberg Lab 916-752-9021 Fax 916-752-4604 dlholmberg-at-peseta.ucdavis.edu
Dear All, We are are multi-user electron microscope facility which has an extensive range of equipment and uses a wide range of techniques. Five technical staff from three contributing University departments are employed full-time to undertake for work coming from both inside and outside the University.
The role of the 'academic in charge' of the facility is shortly to come up for reassessment and so it is a pertinent time for us to reconsider what that role should entail. We are looking for feedback from other individuals as to what they see the contribution of an academic in this environment should be. Any opinions/suggestions?
} The role of the 'academic in charge' of the facility is shortly to come up } for reassessment and so it is a pertinent time for us to reconsider what } that role should entail. } We are looking for feedback from other individuals as to what they see the } contribution of an academic in this environment should be. Easy - 1. Keep abreast of all techniques which a) you have, b) exist, and c) potentially exist. 2. Be an expert practionioner of two or more of these. Publish a lot in your own name. 3. Give advice on the application of all techniques and on the high-level interpretation of all results. Publish jointly with others. 4. Raise funds to replace equipment and to buy new techniques as they become applicable. 5. Make sure all users publish, and tell you about it! 6. Establish a reputation as a scientist in some major subject area, not just as a microscopist. Publish a lot. 7. Keep friendly with the heads (or budget controllers) of all potential user departments. 8. Get to know lots of people in your institution by playing sport/drinking/etc with them. 9. In your spare time, publish some more.
I offer this advice after 20 years of running such a facility!
Peter Goodhew
---------------------------------------------------------------------------------------------------------- Professor Peter J Goodhew, Department of Materials Science & Engineering University of Liverpool LIVERPOOL Fax (44) (0)51 794 4675 L69 3BX, UK Tel (44) (0)51 794 4665 (secretary Debra) ---------------------------------------------------------------------------------------------------------- inter alia: Director of the MATTER project for educational software ----------------------------------------------------------------------------------------------------------
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Institute of Physics Publishing Techno House Tel: +44 272 297481 Redcliffe Way Fax: +44 272 294318 Bristol BS1 6NX England Telex: 449149 INSTP G
I am the Publisher of the journal Bioimaging which I hope you have heard of. I would like to place information about the journal, including tables of contents, on your bulletin board. Will this be possible and how should I do it?
Several years ago there appeared an article in Science discussing laboratory space design and a recent "New" model being implemented in England. Does anyone remember this article, and what has happened to the English experiment?
Microscopy List {microscopy-at-aaem.amc.anl.gov} , Functional Neuroimaging List {lat-at-po.cwru.edu} , Confocal Microscopy List {confocal-at-ubvm.bitnet} , Cell Bio List {cellbiol-at-net.bio.net} Message-Id: {Pine.3.05.1.9502081015.B14211-a100000-at-pauline.sdsc.edu} Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII
Can Anyone point me to sources (free to minimal cost) of analog (VCR tape) or digital timelapse movies of cells in culture? This is not for commercial use, only presentation demonstration. Of course I would credit each source in the presentation. Thanks in advance for any help you can give.
Marc
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
Dr. Neal D. Evans Shared Research Equipment Program Oak Ridge National Laboratory Building 5500, MS 6376, Oak Ridge, TN 37831-6376 voice(615-576-4427) fax(615-574-0641) email(evansnd-at-ornl.gov
Is anyone aware of a company that sells reconditioned SEMs?
+---------------------------------------------------------+ | Stuart Wright Los Alamos National Lab, MST-6 | |.........................................................| | Mail Stop G770 phone: (505) 665-3647 | | Los Alamos, NM 87545 fax: (505) 667-5268 | +---------------------------------------------------------+
------ Forwarded message ends here ------
******************************************************************* Daniel E. Sampson dsampson-at-earthsci.ucsc.edu Instrumentation Specialist Phone: (408) 459-4992 Earth Sciences FAX: (408) 459-3074 University of California Santa Cruz, CA 95064 *******************************************************************
Stuart, Have you tried contacting the instrument manufacturers themslves? I believe that JEOL and Hitachi, for instance, may sell the reconditioned SEMs that comes in on trade-ins. Failing that, give us a call, we'll try to direct you to more sources.
Ellie Solit, Publisher of MICROSCOPE TECHNOLOGY & NEWS AND THE MICROSCOPE BOOK
On Wed, 8 Feb 1995, Stuart Wright wrote:
} Is anyone aware of a company that sells reconditioned SEMs? } } } +---------------------------------------------------------+ } | Stuart Wright Los Alamos National Lab, MST-6 | } |.........................................................| } | Mail Stop G770 phone: (505) 665-3647 | } | Los Alamos, NM 87545 fax: (505) 667-5268 | } +---------------------------------------------------------+ } }
Dear All, Further to my message of 8.2.95, thank you very much to those who have responded so candidly to my request for people's feelings and experiences on this topic. I appreciate that it can be a sensitive issue, not helped when one is replying to someone who doesn't even identify themselves properly. As I neglected to state who I am and where I work (I changed computers days ago and forgot to put the automatic signature on - the ramifications of this I am just becoming aware of...) that info now follows. Maybe now I'll get some more replies...
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
I have been asked to do particle sizing of a 10% fat immulsion (pH 8.0) and had hoped to be able to do a very quick negative staining procedure as this may need to be run frequently. I toyed with the idea of freeze fracture, but the time involved is not convenient for lots of runs. Has anyone tried this? What stains would you suggest? Is Osmium vaper fixation nessecary?
We have considerable experience with negative stain of phospholipid vesicles, lipoproteins, and triacylglyceride emulsions. In these circumstances fixation is not critical, and in fact can produce artefacts. PTA stain seems to work best. We concentrate our sample on grid by drying multiple drops under gentle nitrogen gas stream prior to negative stain. This avoids clumping in the suspension. Some sizing artefacts can occur as dryed particles of large size can deform slightly. See Ganz et al, 1991, J Lipid Res 31:163 for nice discussion of this. Good luck-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Thu, 9 Feb 1995, Kathy Walters wrote:
} Dear Fellow Microscopists, } } I have been asked to do particle sizing of a 10% fat immulsion (pH 8.0) } and had hoped to be able to do a very quick negative staining procedure as } this may need to be run frequently. I toyed with the idea of freeze } fracture, but the time involved is not convenient for lots of runs. Has } anyone tried this? What stains would you suggest? Is Osmium } vaper fixation nessecary? } } Thanks for any help! } Kathy Walters } } }
Dear Kathy, Staining is not really my field, but I'd suggest a water-soluble heavy metal and NO osmium. The Os would only dissolve in the lipid and reduce con- trast. If possible, looking at a frozen, hydrated (or lyophyllized in-situ) specimen would be best. You don't specify either the matrix of the emulsion (I assume aqueous) nor the technique to be used (I assume TEM), but a possible protocol would be to add the stain to the emulsion and rapidly freeze, then cryo-section, examine on a Friday, raise the temp to ~-90C, come in Saturday and raise the temp to -80C, Sunday to -70C, and look at the freeze-dried spec- imen Monday. If you can do the particle sizing from the frozen-hydrated spec- imen, the last steps can be omitted. Keeping the stain strictly in the aqueous phase should prevent size changes, etc. in the lipid drops. Good luck. Yours, Bill Tivol
E.J. Fjeld Co, 3 Executive Park Drive North Billerica MA 01862 Phone 508-667-1416
Provides reconditioned AMRAY microscopes. They also make special stages and accessories. They've been around for a long time and are reliable.
Jacob Bastacky, MD 1-116 Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Stuart, my attempts to reply to your question by email have resulted in 6 notices of undelivered mail. I left a voice message on your phone this morning. I think we can help you in your search.
Regards, Ellie Solit, Publisher/Executive Editor of Microscope Technology & News and The Microscope Book, a Smart catalog.
On Thu, 9 Feb 1995, Stuart Wright wrote:
} Is anyone aware of a company that sells reconditioned SEMs? } } } } +---------------------------------------------------------+ } | Stuart Wright Los Alamos National Lab, MST-6 | } |.........................................................| } | Mail Stop G770 phone: (505) 665-3647 | } | Los Alamos, NM 87545 fax: (505) 667-5268 | } +---------------------------------------------------------+ } }
Message-ID: {95020916094142E.RQDA-at-USCN.USCN.UGA.EDU} (UMass-Mailer 4.04) Neuroscience List {neur-sci-at-net.bio.net} , Microscopy List {microscopy-at-aaem.amc.anl.gov} , Digital Video List {digvid-l-at-ucdavis.edu} , Confocal Microscopy List {confocal-at-ubvm.bitnet} , Cell Bio List {cellbiol-at-net.bio.net} Message-Id: {Pine.3.05.1.9502091131.A22203-e100000-at-pauline.sdsc.edu} Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII
I thought this post should be quickly disseminated.
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
---------- Forwarded message ----------
THE FOLLOWING RELEASE MOVED OVER PR NEWSWIRE ON TUESDAY, FEBRUARY 7, 1995 AT 11:42 AM, PST.
Apple Announces QuickTime Conferencing Open, Cross Platform Conferencing, Collaboration and Multimedia Communications Technology
SAN FRANCISCO, California--February 7, 1995--Apple Computer today announced a cross platform conferencing, collaboration and multimedia communications technology that allows personal computer users to share real-time information, images and sound anywhere in the world. Apple is currently making the technology, called QuickTime Conferencing, available to corporate allies who plan to create or have announced they are creating end user applications based on the technology. QuickTime Conferencing is a standards- based architecture that allows users to:
-- video conference and collaborate--to share and annotate text, images, screen capture, sound, video and virtual scenes real-time among fellow conference participants in a variety of locations worldwide. QuickTime Conferencing allows users to record conversations and transform those conversations into QuickTime movies. All of this can be done on a variety of networks such as an Integrated Services Digital Network (ISDN), the worldwide internet, local area and wide area networks and Asynchronous Transfer Mode (ATM) networks. QuickTime Conferencing can be used by a number of simultaneous users, the total number being only by available network bandwidth.
-- conduct cross platform video conferencing connectivity between Macintosh computers, PCs, UNIX systems and room-based conferencing systems through the use of the H.320 worldwide teleconferencing standard.
-- broadcast and view multimedia content--digital audio, music and video on a local or wide area network.
Through alliances QuickTime Conferencing technology is expected to yield product bundles such as: -- Apple Media Conference Kit--Consisting of the QuickTime Conferencing system extension, the Apple Media Conference application and a high quality, color video camera. -- Apple Media Conference Pro Kit--Consisting of the QuickTime Conferencing system extension, the Apple Media Conference application, a color video camera and an H.320 codec/ISDN adapter board. Being developed by Sagem/SAT, a leading international communications product company, the board is designed to allow interoperability between platforms (Power Macintosh to Macintosh, PC, UNIX and room systems) and full-screen image sharing. --Complete Media Conference System--Consisting of an Apple Media Conference Kit, a Power Macintosh 7100 AV, a 17 inch color monitor, external speakers and a keyboard.
Because QuickTime Conferencing is software-based, it is easily incorporated into new and existing third party products. As such, Apple believes that QuickTime-compatible products could yield extremely affordable prices: -- Apple Media Conference Kit--under $200 -- Apple Media Conference Pro Kit--under $1,750 -- Complete Media Conferencing System--under $6,000
Apple is working with a wide range of companies including telcos, network, software and hardware providers and developers to provide a range of solutions that take advantage of the benefits of QuickTime Conferencing (see associated releases). These allies have announced that they expect to make products available in the second quarter of 1995. From the home office to university campuses to the multinational enterprise network, QuickTime Conferencing will allow users to communicate with people across the country or across the world. Users won't have to worry about whether their hardware equipment, networking equipment and applications are compatible with the solutions being used on the other end of the network line. QuickTime Conferencing is designed to be fully operational with H.320 standards-based systems. "The introduction of QuickTime Conferencing will not only extend Apple's leadership in multimedia, but will make an important difference in the video conferencing and collaboration market," said Rick Shriner, vice president of Apple's Core Technologies Group. "Our goal in designing QuickTime Conferencing was to develop a solution that allowed people the opportunity to communicate and collaborate. By making it open in every sense of the word, our users can metaphorically break down the walls of their homes, schools and offices and expand the boundaries of their lives." QuickTime Conferencing users can have access to people, information, sights and sounds that could never be combined before. For example: -- An author in Tokyo, Japan and her publisher in San Francisco, California can view and discuss cover art for a new novel. They can each view the design at several different angles, change the visual perspective of the artwork, and annotate the image and accompanying text for the other to see. -- A sixth grade class in Dallas, Texas can discuss and view the effects of global warming with an environmental scientist at U.C. Berkeley's Lawrence Labs in California by using QuickTime Conferencing over the internet. -- A special effects producer in Hollywood, California can take a movie director on the East Coast through a virtual tour of a proposed set design. While the producer records their discussion as a QuickTime movie, the director can pan around the scene, zoom in to look at props and view the set design from a variety of angles. -- A breast cancer patient and her doctor in Fargo, North Dakota can consult with a leading oncologist in Boston, Massachusetts on her prognosis and course of treatment. The Boston physician can view her mammograms and annotate her medical chart as they converse. -- A CEO's company-wide address can be broadcast for easy viewing by all employees at their personal desktop.
Because QuickTime Conferencing allows for sharing of multimedia data and reduces the time and expense of travel, it allows people to be more productive than ever before. "In the past people found video conferencing easy to resist because prices were high and the number of people they could communicate with was extremely limited," said Rick LeFaivre, senior vice president of the Apple Technology Group. "Now for what we expect to be very aggressive prices, people can conduct a media conference with virtually anyone, anywhere in the world. A Power Macintosh QuickTime Conferencing user can share QuickTime VR (virtual reality) images, annotate text documents and share digital music over networks from basic rate ISDN to the internet to ATM." Because QuickTime Conferencing is a software-based architecture, application developers, communications providers and hardware vendors can easily develop compatible solutions. For example, Crosswise Corporation, the maker of Face to Face, a cross-platform document conferencing application, developed a QuickTime Conferencing-compatible version of their software in just one month. A QuickTime Conferencing compatible application shares the interface of other QuickTime Conferencing-enabled third party applications, so customers can begin using applications quickly and easily. QuickTime Conferencing is based on Apple's award winning QuickTime technology. It is a conferencing architecture which allows support for both industry standards such as H.320, as well as proprietary architectures, and codecs such as Indeo by Intel Corporation. QuickTime Conferencing is transport, compression and media-device independent. Apple's built-in AV capabilities combined with the performance of the PowerPC RISC architecture, make it easy for users to make multimedia connections with others on the information superhighway almost as soon as they pull QuickTime Conferencing out of the box. "Having QuickTime Conferencing available in my home, office, and studio literally allows me to be in multiple locations at one time--it's the next best thing to having a Star Trek transporter," said Los Angeles-based screenwriter and multimedia special effects consultant Michael Backes, co-author of the screenplay for Jurassic Park and other motion pictures. "Within the next few months, I'll be counting on QuickTime Conferencing as the backbone for my business." "The short and sweet of QuickTime Conferencing is that it requires less network bandwidth and uses innovative technology," says Matt Ghourdjian, National Director of Technology at Howrey & Simon, a 300-lawyer law firm serving Fortune 50 clients. Howrey & Simon intends to use the product to send QuickTime movies of depositions and re-enactments for lawyers to use in court; for live document sharing; for consultation between partners; and to conduct tours of the firm's Washington, DC office from Los Angeles. "It's simply outstanding," says Chris Masten, Howrey & Simon's Technical Litigation Support Manager. To use the Apple Media Conference Kit on the Macintosh, users need at least 16 Megabytes of RAM, a 68040 or PowerPC-based Macintosh, System 7.5, a network interface such as Ethernet, ISDN, Token Ring, and optionally the ability to digitize audio and video using the built-in AV subsystem or a third party digitizer card. To use the Apple Media Conference Pro Kit on Macintosh, users need at least 16 Megabytes of RAM, an AV PowerPC-based Macintosh and an ISDN connection. To communicate with QuickTime Conferencing users from the PC and other platforms, users will need an H.320 compatible codec on their machine, available from a variety of vendors. QuickTime Conferencing technology is currently under development and products using the technology have not yet been completed. Apple will provide pricing and availability information when products are completed and ready for release. Apple Computer, Inc., a recognized pioneer and innovator in the information industry, creates powerful solutions based on easy to use personal computers, servers, peripherals, software, online services, and personal digital assistants. Headquartered in Cupertino, California, Apple (NASDAQ:AAPL) develops, manufactures, licenses and markets products, technologies and services for the business, education, consumer, scientific & engineering and government markets in over 140 countries.
-30-
Apple, the Apple logo, QuickTime and Macintosh are registered trademarks and Power Macintosh is a trademark of Apple Computer, Inc. Additional company and product names may be trademarks or registered trademarks of the individual companies and are respectfully acknowledged.
END
Applelink pathway: News Break Apple & Industry News PR Express Apple Press Releases 2/7/95
I know of a company that reconditions old AMRAY SEMs. The owner used to work at AMRAY before starting his own company. Company is E.J. Feld located in Massachusetts. I can give you the phone on Monday, 2/13 when I return to work. Dr. David A. Shifler Powell Labs Ltd. Baltimore, MD 31231 (410) 327-3500 (410) 327-7506 (FAX)
Thanks to all the replies to my recent TEM survey, I am well on my way to preparing the survey for publication. I need some information -- I'm a chemist, not a microscopist -- What do TEM users see as important trends in TEM? New technology? Important applications? Is there other technology that is replacing TEM in any way. How important are accessories like EDS and EELS? For what uses?
Any comments on today's TEM would be greatly appreciated -- I'll send a copy of the survey to any contributor.
Does anyone in the group have the phone number for PRESENTATION TECHNOLOGY? We are in the information gathering stage for the purchase of a 35mm slide maker and the phone number we have (408-774-3733) is not correct.
Thanks for your help.
Doug
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (602)626-2824 dcromey-at-ccit.arizona.edu
} Dear Microscopists, } } Does anyone know if there is any recent books about applications of } microbeam techniques to mineralogy and petrology? } } The one I have was published by Mineralogical Association of Canada } (Short Course in Microbeam Techniques) in May 1976. } } Thanks in advance. } } Long LIang } ARCO EPMA/SEM Lab } PLano, TX
Try: McLaren, A.C., 1991, TEM of minerals and rocks, Cambridge University Press, Cambridge
Boland, J.N. & FitzGerald, J.D. (eds), 1993, Defects and processes in the solid state: geoscience applications, Developments in Petrology, Vol 14, Elsevier, Amsterdam
Buseck, P.R. (ed), 1992, Minerals and reactions at the atomic scale: TEM, Mineralogical Society of America, Reviews in Mineralogy, vol 27.
Cheers Timon
------------------------------------------------ Timon Fliervoet, Timonf-at-earth.ruu.nl, Faculty of Earth Sciences, Department of Geology, Utrecht University, P.O. Box 80.021 3508 TA Utrecht, the Netherlands. tel: ++31 30 - 535054, fax: ++31 30 - 537725
Help please!! SiSTek is looking for TEM analytical services in the southwestern US, preferably in the Phoenix metropolitan area where we are located. We are a company that provides consultant services to a number of manufacturers of Si-related deposition systems and need *local* (turnaround time and iterative analysis is important for our clients) TEM support. We believe there is a company in the Phoenix area offering TEM services, but can't find the name. Does anyone know the name/contact?
Subject: Time: 9:13 AM OFFICE MEMO Ecomet Polisher for sale Date: 2/14/95
FOR SALE: ECOMET 1 8" Polisher/grinder, complete with various polishing discs, alumina and lapping oil. 115 volt, 5 amp, new price in 1988 = $1250.00 Best offer. Call Doug Davis of EM Lab at UC Berkeley at (510) 642-2085 or e-mail: doug_davis-at-maillink.berkeley.edu
Any tips on cutting wells out of 24 well cell culture plates would be greatly appreciated.
Joe Joseph A. DeMaro Washington University Medical School Department of Neurology 660 S. Euclid Rm 212 Biotech St. Louis, MO 63110 jad1-at-cec.wustl.edu 314-362-9448
LaserMaster Corporation - Imaging Division is the ONLY company that manufactures a 1800 dpi plain paper laser printers. I work for LaserMaster and have just completed printing the 100/300 dpi Round Robin Greyscale Test images. If you are interested in obtaining them, you may e:mail me at Gregb-at-Sales.LMT.com and I will mail you a printout of the test images from the LaserMaster printer. I can be reached at 1-800-950-6363 Ext: 3207 if you have questions about your specific situation and resolution needs.
Thank You all, Greg Begin - LaserMaster Corp. Scientific/Medical Imaging Div. /\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\\/\/\/\/\/\/\/\/\/
} Date sent: Thu, 09 Feb 95 07:42:30 -600 } From: Supratik_Guha-at-mail.mmmg.com (SG) } To: microscopy-at-aaem.amc.anl.gov } Subject: printers for gray scale prints
} I am looking around for a gray scale printer to attach with our Gatan } slow scan CCD image aquisition system and would appreciate any } suggestions. We would prefer not to get a dye-sub printer due to the } high costs of printing. I understand that there are 1800 dpi laser } printers available, could someone point out manufacturers for these } machines? } } Supratik Guha } Senior Materials Scientist } 3M Corporate Research Labs. }
When I want to process cells for SEM from 24 well culture dishes I use a Bunsen burner and a scoopula (the curved metal device for scooping out dry chemicals). First, the cells are fixed as usual with glutaraldehyde then washed in buffer and distilled water. Then, working within a fume hood and wearing a heatproof glove, the scoopula is heated until red then touched to the underside of the culture dish. The curved metal is about the right size for the 24 well dishes. It takes 2 or 3 times to heat and cut until the whole bottom is released. Once the initial cut is made you need to keep the cells wet and this is easily done using a squirt bottle of distilled water. This technique does not appear to cause damage to the cells but we normally look at the more centrally positioned cells to avoid any artefacts. Hope this helps.
Nancy Smith Cal State Hayward nsmith-at-csuhayward.edu
Hello, everyone: Does anyone know where I can find some reference on measurement of electron mean free path of different materials? The mean free path I mean here is the mean free path for measuring the thickness when doing EELS, so this includes electrons of all the energy losses rather than one particular energy loss. We are particularly interested in getting the right electron mean free path for Chrome Oxide and evaporated Carbon. Thanks a lot.
Eric Wang FB-10 Roberts Hall Univ. of Washington Seattle. Wa 98195 (206) 543-1514
} Any tips on cutting wells out of 24 well cell culture plates would be greatly } } appreciated. } Joe } Joseph A. DeMaro
Depending on what exactly it is you are doing, how about using Thermanox (Thermonox?) plastic slides on the bottom of the wells - they make them specially to fit into the wells of 12 well plates and possibly the 24 well ones too. They come sterilised and you just pop them into the well before you add medium and cells and remove later, fix, dry etc and mount on a stub. The slide surface properties are supposed to duplicate the ordinary well bottoms. Its easier than cutting wells out of the bottom of the trays. Regards R Easingwood
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
I work for an OEM of scanning electron microscopes and am interested in keeping up with the latest technology and issues. I would like to subscribe to the mailing list. Could you please let me know what I need to do.
We process a considerable number of cell cultures for both SEM and TEM, and have found what we consider to be the ideal system. For this purpose, we have our investigators culture their cells in Leighton tubes...these are cell culture tubes with a flat bottom which holds a long, narrow plastic coverslip. Once the cells are attached, the medium is replaced with fixative, then the coverslip (which is attached to a nifty little handle) is removed. The plastic on the coverslip is impervious to all solvents used in microscopy, and can be embedded and sectioned for TEM.
I wouldn't think of doing it any other way.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
We have two Beseler enlarger needing adjustments. Any information on available service person in the south, please remit details directly to me. Number I called was disconnected!
************************************************************ *Cesar D. Fermin, Ph.D \|*|/ Fax (504) 587-7389 * *Tulane Medical School /|*|\ Answ. Mach.(504) 584-2618 * *Pathology/SL79 \|*|/ Secretary (504) 584-2436 * *New Orleans, La 70 112 /|*|\ Lab (504) 5841 * *Fermin-at-TMC.Tulane.edu -} Director of Morphological Services* ************************************************************
Back around '82 or '83 I found that I had a problem with my Besseler's constantly slipping focus just a tiny bit. So I put pipe clamps (those metal rings that can be tightened of loosened with the turn a a screw) on the metal guides for the focus which I would tighten when focusing the bellows particulalry tight. -mc
On 15 Feb 1995, Fermin, Cesar wrote: } ref.: Beseler enlarger tune-up.
For those interested, Leighton tubes for cell culture are manufactured by Corning Costar and are available from Fisher Scientific (Cat.# 07-200- 367). They appear in the 95-96 catalog on page 721.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
Dear Eric, If no expert in the field has a table of the mfp's, I can fax you tables of stopping powers for electrons in carbon, oxygen, iron & titanium-- you have to interpolate for chromium and calculate the mpf from dE/dx. Good luck. Yours, Bill Tivol
Many thanks to all who responded for my call for help with locating TEM service near us in the Phoenix metro area. As we thought, there is an established group in Phoenix who have been around for a few years and who provide TEM services.
For anyone else who might be interested, the company is called NanoTEM, Inc, 7620 E. McKellips Rd., Suite 4109, Scottsdale, Arizona 85257, phone 602 759 2808, fax 602 947 7615.
Greetings, Is there something out there that will make a thin film that isn't formvar? I have been using wire loops coated with a film of 1.2% formvar to support my small samples during rapid freezing and substitution. This works fine for acetone substitution but we would like to try Tetrahydrafuran (THF) as a substitution medium and, alas, THF eats the formvar.
We get a formvar film on the wire loop by casting small rectangles of formvar on water and then trasfering one to a loop.
Are there other compounds that could be used to make a film across the loop and that might survive THF??
I am trying to prepare polished sections from steel samples for EPMA analysis. Are there any recommended abrasive/size for rough grinding, fine grinding, rough polishing, and final polishing?
Your help is high appreciated.
Long Liang ARCO EPMA/SEM Lab Plano, TX LLIANG-at-is.arco.com
} } I am trying to prepare polished sections from steel samples for EPMA } analysis. Are there any recommended abrasive/size for rough grinding, } fine grinding, rough polishing, and final polishing?
I have always stuck to the simple silicon carbide paper (in steps from 120 to 1200 grade) and then diamond (6,3,1um) route, and found no problems with that.
Doug
+------------------------------------+ | Dr Douglas Arrell | | Mechanical Performance and Joining | | Institute for Advanced Materials | | 1755 ZG Petten | | Netherlands | | {ARRELL-at-JRC.NL} | | Tel. (+31) 2246 5287 | | Fax (+31) 2246 1917 | +------------------------------------+
CALL FOR PAPERS STUDENT COMPETITION TO BE HELD AT THE UNIVERSITY OF WISCONSIN WHITEWATER, WISCONSIN Friday, March 24, 1994 AWARDS: FIRST PLACE $100 SECOND PLACE $75 THIRD PLACE $25 Abstracts will be published in Midwest Microscopy. Microsgraphs from first place winner will be on the cover of Midwest Microscopy. Students will be judged on written abstract, presentation, and quality of study. Student competition is open to undergraduate and graduate students and may involve any type of microscopy. Student and sponsoring faculty member must be members of MSEM. Abstracts should be submitted before March 15 to : Dr. Lance Urven University of Wisconsin- Whitewater 800 West Main,Whitewater, WI 53190
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu Time: 11:59 AM
Date:2/20/95 URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html NC EMAL
I have some colleagues that want to electropolish some fairly heavily deformed steel. Composition is: 0.4% C 0.6-0.9%Si 22-24%Cr 7-9%Ni Trace N Balance Fe. Does anyone have a good starting solution and condidtions for this alloy? Many Thanks. John Mansfield.
} I am trying to prepare polished sections from steel samples for EPMA } analysis. Are there any recommended abrasive/size for rough grinding, } fine grinding, rough polishing, and final polishing?
A step that may be advantageous is the final polish of the steel by use of a colloidal silica type solution [0.05 micron, with 9.8pH]. Dampen the cloth [such as a Buehler Mastertex] with distilled water; apply liberal amount of the solution [such as Buehler Mastermet] to the cloth, and apply firm pressure to soft pressure over a period of ~45 seconds, rotating the sample quite abit. A final word of caution: this solution [Mastermet], besides having the high pH [rough on skin], will crystallize into small, -very hard- particles. Is therefore highly advised to filter the solution a few times into a smaller bottle before each use. Have found this stuff to be -very- effective tho'. LECO also has a similar solution, but have not used it enough to get comfortable with it as the Mastermet.
In the previous steps I have used the 120 to 800 grit SiC papers, then a 9, 6, 1, & sometimes 1/4 micron diamond paste, on Texmet for the rough polish (9 & 6), and then Mastertex for the 1/4u. All depends on the grade and condition of the steel tho'... [all these recommendations are based on me hand polishing individual samples, and 3-6 samples in a ring held in hand]
Hope this helps, -Rob PS: I have no ties with Buehler, just a satisfied customer for +10 years. ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /\v/\ | | Metallographic Lab. | Missouri Speleological Survey /\v/\ | | Rolla Research Center | Bat Conservation International /\v/\ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-rorc.usbm.gov | National Speleological Society #32993 /\v/\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
The techniques described by Rob Tayloe are certainly in agreement with the references I have for polishing steel and will work quite well. The only note I would make is that we do supply a Colloidal Silica which DOES NOT CRYSTALLIZE. This is an important feature as Rob has mentioned because the crystallized particles can be a real problem if you do not realize they are there.
Our Non-Crystallizing Colloidal Silica is available as follows:
Part No. Description Price CS1-16 Non-Crystallizing Colloidal Silica 16 oz bottle $16.00 CS1-128 " " " 1 gallon bottle 78.00
If you'd like more information (MSDS etc) on this prodcut or any of our other products, please let me know.
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
one of my users is looking at TEM of insect heart. Does anyone have some good references on insect heart ultrastructure? Some of the cells associated with the heart appear highly vesiculated at the cell periphery. The morphology of these cells looks good, so we don't feel these stuctures are an artifact, but we have so far been unable to identify what kind of cell or cell type it might be. Can anyone suggest a reference or an investigator we could contact in this regard?
---------------------------------------------------------- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego CA 92182-0057 phone: (619) 594-4523 fax: (619) 594-5676 email to sbarlow-at-sunstroke.sdsu.edu
} SUSAN R. SESACK wrote: } } Would someone please provide me with instructions for enrolling in } the Internet bulletin board for histology and microscopy? Much } obliged.
To subscribe, please send a mail to "Listserver-at-AAEM.AMC.ANL.GOV" with "Subscribe Microscopy your_id-at-e-mail" in the body. Enjoy it.
Holland Cheng -------------------- Structural Biology Department of Biological Sciences Purdue University W. Lafayette, IN 47907-1101
} SUSAN R. SESACK wrote: } } Would someone please provide me with instructions for enrolling in } the Internet bulletin board for histology and microscopy? Much } obliged.
To subscribe, please send a mail to "Listserver-at-AAEM.AMC.ANL.GOV" with "Subscribe Microscopy your_id-at-e-mail" in the body. Enjoy it.
Btw, can someone in the server fix the returning route so that the reply can a global one? I would like to be on a list that I can see discussions (questions and answers) rather than a collection of of questions. Thanks in advance!
Holland Cheng -------------------- Structural Biology Department of Biological Sciences Purdue University W. Lafayette, IN 47907-1101
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ ++++I would like to add a small point about polishing specimens for analysis in the electron probe microanalyzer (EPMA). I prefer to leave the scratches in from the 1/4micron diamond polishing step in order to have an image feature on which to focus with the light optics. Since quantitative x-ray microanalysis samples should be flat-polished but unetched, it is hard to find a suitable surface feature to use for focusing without these fine scratches. For some reading on this, try Chapter 11 of Goldstein et al., Scanning Electron Microscopy and X-ray Microanalysis, 2nd edition, Plenum Press, 1992.
Good luck, Prof. C E Lyman
Electron Optics Lab Lehigh University 5 East Packer Avenue Bethlehem, PA 18015 ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
} } I am trying to prepare polished sections from steel samples for EPMA } } analysis. Are there any recommended abrasive/size for rough grinding, } } fine grinding, rough polishing, and final polishing? } } A step that may be advantageous is the final polish of the steel by } use of a colloidal silica type solution [0.05 micron, with 9.8pH]. } Dampen the cloth [such as a Buehler Mastertex] with distilled water; } apply liberal amount of the solution [such as Buehler Mastermet] to the } cloth, and apply firm pressure to soft pressure over a period of ~45 } seconds, rotating the sample quite abit. A final word of caution: } this solution [Mastermet], besides having the high pH [rough on skin], } will crystallize into small, -very hard- particles. Is therefore highly } advised to filter the solution a few times into a smaller bottle before } each use. Have found this stuff to be -very- effective tho'. LECO also } has a similar solution, but have not used it enough to get comfortable } with it as the Mastermet. } } In the previous steps I have used the 120 to 800 grit SiC papers, then } a 9, 6, 1, & sometimes 1/4 micron diamond paste, on Texmet for the } rough polish (9 & 6), and then Mastertex for the 1/4u. All depends on } the grade and condition of the steel tho'... [all these recommendations } are based on me hand polishing individual samples, and 3-6 samples in a } ring held in hand]
Dear Tobias, A carbon film--evaporated onto freshly-cleaved mica--should do the trick. After evaporation, float the film onto water and pick it up with the loop. You may have to experiment with thickness etc. to get the proper mech- anical strength, but it should certainly survive the THF. You may also want to try a mesh grid instead of the loop if the carbon film is not strong enough. Good luck. Yours, Bill Tivol
Message-Id: {9502212303.AA26486-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Re: TEM/formvar substitute/thin films Orig-Author: {tivol-at-tethys.ph.albany.edu}:ddn:wpafb ----------------------------------------------------------- Dear Tobias, A carbon film--evaporated onto freshly-cleaved mica--should do the trick. After evaporation, float the film onto water and pick it up with the loop. You may have to experiment with thickness etc. to get the proper mech- anical strength, but it should certainly survive the THF. You may also want to try a mesh grid instead of the loop if the carbon film is not strong enough. Good luck. Yours, Bill Tiv
I've done this quickly with "Image" by measuring the distance from the graph axis (x or y) to the data points in question. To calibrate (pixels to data units), just measure the distance between axis values.
On Wed, 22 H Feb 1995 Noonan_Eddie/perth-at-perth.atd.cra.com.au wrote:
} I have been asked by a colleague of mine who at present does not have } access to the Microscopy Listserver and who presently is sourcing a } new SEM for his lab to put out the following question. } } "In the next few weeks I shall be ordering a new analytical SEM. We } will use this SEM almost exclusively for EDS analysis. A motorised } stage will also be required for the SEM to accommodate overnight runs. } Therefore I require an ultra stable, reliable instrument. If any one } with experience in this area has any thoughts I would be grateful to } hear from you." } } Thanks in advance } } Mark Stewart } } Replies to: ejn-at-perth.atd.cra.com.au }
Jan Coetzee and Philip Oshel have posted about blood fixation but I havn't seen any replies on this topic. We have a project that will try to tie distortion of red cells to heat stress so fixation needs to be as artefact free as we can get it. Please Jan or Philip can you mail me with the best method you can recommend?
1) In the February '95 Biotechniques, New Products section, there's a blurb for a "Probe Clip" single slide incubation chamber sold by Grace Bio-Labs. Anyone have experience with these and/or a contact phone/fax # for Grace Bio-Labs?
2) A grad student in our lab has been doing ImmunoGold Silver (LM) immunostaining followed by BrDU - HRP - DAB for a second antigen. The silver was originally present, but faded out in the second reaction. Could have resulted from a number of factors, but what we were wondering is - is it possible to stabilize the silver with a sodium thiosulfate "fixer" step after the IGSS to protect it in subsequent immunoreactions, dehydrating and coverslipping? Any practical suggestions would be appre- ciated.
3) A colleague in Australia wants to purchase an antiserum to met- enkephalin. We have been buying from Incstar and getting good results, but the Australian distributor for Incstar charges outrageous prices. Does anyone know of an alternate antibody supplier that may be less ex- pensive for Australian customers?
------------------------------------------------------------------ |Glenn Holm *mime mail ok* Internet:karuzis-at-wccf.mit.edu | |M.I.T Dept. of Brain + Cog. Sci. This VAX doesn't do NeXTmail | |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | ------------------------------------------------------------------
Recently I posted a request for methods for lipid sizing. I have put together a summary of that request. If anyone would like a copy I will be happy to send you one, but it is rather lengthy for the listserver.
Marc C. Brande, M.S. SD3D Email List:3D Imaging San Diego 3D Imaging Group To subscribe/unsubscribe:listserv-at-bobcat.etsu.edu 3840 Camino Lindo To post a message:sd3d-at-bobcat.etsu.edu San Diego, CA 92122 Email:BRANDE-at-SDSC.EDU Voice:619-587-4830
On Thu, 23 Feb 1995, Glenn Holm wrote:
} 3 separate requests for information: } } 1) In the February '95 Biotechniques, New Products section, there's } a blurb for a "Probe Clip" single slide incubation chamber sold by } Grace Bio-Labs. Anyone have experience with these and/or a contact } phone/fax # for Grace Bio-Labs? } } 2) A grad student in our lab has been doing ImmunoGold Silver (LM) } immunostaining followed by BrDU - HRP - DAB for a second antigen. } The silver was originally present, but faded out in the second reaction. } Could have resulted from a number of factors, but what we were wondering } is - is it possible to stabilize the silver with a sodium thiosulfate } "fixer" step after the IGSS to protect it in subsequent immunoreactions, } dehydrating and coverslipping? Any practical suggestions would be appre- } ciated. } } 3) A colleague in Australia wants to purchase an antiserum to met- } enkephalin. We have been buying from Incstar and getting good results, } but the Australian distributor for Incstar charges outrageous prices. } Does anyone know of an alternate antibody supplier that may be less ex- } pensive for Australian customers? } } ------------------------------------------------------------------ } |Glenn Holm *mime mail ok* Internet:karuzis-at-wccf.mit.edu | } |M.I.T Dept. of Brain + Cog. Sci. This VAX doesn't do NeXTmail | } |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | } ------------------------------------------------------------------ }
I believe this question was asked on this list before, so I am sorry if I am repeating. A colleague here is having problems with negatives being ruined by static discharge in a JEOL 2010 TEM. JEOL suggests drying the negatives inside the TEM vacuum which works but is really not good and very inefficient. The problem began only recently when atmospheric humidity is increasing here. Does anyone know the cause of the static discharge problem and/or have a practical solution.
Norman Elliott | E-mail: nee-at-lanl.gov Los Alamos National Lab | Fax: 505-665-2104 MST-7 MS E549 | Voice: 505-667-1587 Los Alamos, NM 87545 |
I would like to announce the establishment of an FTP site for the Macintosh/Power PC based digital imaging microscopy software IPLabspectrum from Signal Analytics Corporation. There are several reasons for development of this site: 1. To provide individuals interested in developing a digital imaging microscopy system a chance to access and download demonstration versions of this particular Macintosh/PowerPC based digital imaging software for evaluation purposes.
2. To provide users of IPLabspectrum software for a site to download and upload user-generated system extensions and scripts .
Although the software is from a commercial source, the establishment of the FTP site was a decision by made by myself and other users of the Digital Imaging Microscopy Facility here at the University of Alabama at Birmingham as one way of promoting the digital imaging microscopy as a research tool. Signal Analytics Corporation, the developers of the software, has generously made available demonstration versions of their entire software line for FTP download purposes. If any other developers of imaging software wish to use this site as a repository for demonstration versions of their software I would be more than happy to accomodate them.
The demonstration versions of this software are organized into several folders, and separated into Macintosh and PowerPC versions of that software. There are versions of the software that support specific imaging boards (e.g. Scion LG-3) in the demo folders. Also in the site are demo versions of calcium ratio (IPLab Ratio), Three Dimension Reconstruction, and Multiprobe (FISH) extensions. In addition is a separate program for reading scanned images of electrophoretic gels. (IP Lab Gel)
There is a folder (directory) for Uploading of Scripts, Extensions, Messages, ect. provided-however the caveat there is that downloading the extensions from that directory is at your own risk, since this is a public access directory. If you choose to download from this directory, please take the time to scan the files for viruses or other nasties. Our aim is to monitor the upload directory as the usage increases, test the material in the directory for problems (bugs, viruses, and other annoying creatures) and then shift those programs that we consider problem free to a specific download-only directory. At present this download-only directory for user-uploaded files does not exist.
To access the site use the address FTP.BMG.UAB.EDU, logon as ANONYMOUS to enter the public directory. In the directory is a subdirectory (folder) IP_Labspectrum. Within this subdirectory are other directories containing the various demos of the software. I also included the shareware programs FETCH and BinHex 5.0 in a directory named FTP_SITE_Utilities for individuals (like myself) who rather use FETCH as a front end for FTP browsing and transfer.
This site is still undergoing development, so please forgive the style of organization. Any comments, suggestions, ect. about the site, digital imaging microscopy would be greatly appreciated. My e-mail address is KJMcCarthy-at-CellBio.BMG.UAB.EDU
Best Regards Kevin McCarthy Assistant Professor Department of Cell Biology University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029
Return-Receipt-To: FRAWLEY-at-a1.resmel.bhp.com.au Registered-Mail-Reply-Requested-By: FRAWLEY-at-a1.resmel.bhp.com.au Mr-Received: by mta VULCAN.MUAS; Relayed; Fri, 24 Feb 1995 11:44:58 +1100 Mr-Received: by mta VULCAN; Relayed; Fri, 24 Feb 1995 11:59:09 +1100 Disclose-Recipients: prohibited
I am interested in using an extraction replica technique to determine MnS precipitate size distributions in a TEM.
The technique being considered is as follows: -Polish sample -Etch to give some surface relief -Carbon coat -Using blade cut 1mm size grid on surface -Release carbon film containing ppt's. from surface using etchant -Wash carbon replica in alcohol and water -Position on TEM grid.
My problem is finding an etchant that will not attack the very small MnS ppts. It has been reported that some etchants attack the Mn in these ppt's.
Any suggestions on the technique or the etchant would be appreciated.
Regards Leo D Frawley frawley-at-a1.resmel.bhp.com.au
} Date sent: Thu, 23 Feb 1995 14:09:10 -0700 } To: microscopy-at-aaem.amc.anl.gov } From: nee-at-beta.lanl.gov (Norman Elliott) } Subject: TEM negatives
} Dear list, } } I believe this question was asked on this list before, so I am } sorry if I am repeating. A colleague here is having problems with } negatives being ruined by static discharge in a JEOL 2010 TEM. JEOL } suggests drying the negatives inside the TEM vacuum which works but is } really not good and very inefficient. The problem began only recently when } atmospheric humidity is increasing here. Does anyone know the cause of the } static discharge problem and/or have a practical solution. } } We had this static discharge problem when first we bought our 2010. It was solved when JEOL replaced the Teflon drive gear (large) in the camera with a conducting (metal) one. Coating the gear with a conducting spray didn't work! We also installed a separate vacuum desiccator for the plates (diffusion pump and liquid nitrogen trap).
Peter Smith ps-at-bunyip.ph.rmit.edu.au RMIT App Physics Ph: (03) 660 3390 Office GPO Box 2476V (03) 660 2205 Lab Melbourne Fax: (03) 660 3837 Vic 3001 AUSTRALIA
About your discharge problem, we believe this is indeed strongly linked with the vacuum. Therefore we always use one or more desicators before the plates enter the microscope. This way also the vacuum in the microscope restores faster. The problem also depends on the plates: we had the experience that Ilford plates always gave discharge, while Kodak and Agfa did not in the same environment. Also handling of the plates can be important: sliding plates over one another can cause discharges before and after use in the microscope. Hope this can be of help, Nick Schryvers
I had some static discharge problems with my JEOL 1200exII some time ago. Every 8th to 10th neg had streaks on it. they were thin and sharp with two or three finger like projections coming fron the same point. The service tech. cleaned the camera (several times) and that cleared the problem up. It did take several tries and return trips to get the dirt (thought to be a metal fragment) cleaned up.
I hope this helps. Larry Hawkey hawkey-at-neuro.duke.edu 919-641-6425
Norman, Although I am not familiar w/ this particular model of JEOL, could it be possible that the static discharge is occuring before or (more likely) after the film has gone into/come out of the scope? We had an individual in the lab I was in before who tended to wear synthetic fabrics and on particularly dry Arizona days they could really ZAP the film. One solution is to wear a wrist grounding strap when handling the film (like the kind that people are supposed to wear when working inside their computers). Hope this helps. Doug
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (602)626-2824 dcromey-at-ccit.arizona.edu
Dear Norman, We, too, have had occasional problems with static discharges on our films--especially with LoDose, which is very sensitive to light. Our worst problems occur when the humidity is low. You must be very careful when sepa- rating films from the stack and when removing the stack from the package. You might also check if your lab coats produce static electricity--we just got new polyester coats which are terrific generators. If you are completely dark-ad- apted and loading the folm in total darkness, you can see the discharges as they occur, so at least you will know what is going on. [oops, I meant film] If your desiccant is too efficient, the discharge problem will be worse. Good luck, sometimes the procedures necessary to prevent these discharges are te- dious. If all else fails, try using 4489 film--it's not nearly as sensitive as SO163, but it is much finer grain and gives excellent images. We've never had discharge problems with it, probably because discharges are not intense enough to show up. Yours, Bill Tivol
In the past we too have had problems with static discharge ruining negatives (the film was SO163 being used in a JEOL 4000EX). We found that the discharges were coming from our vinyl gloves; when we switched to cotton gloves the problem went away. Additionally, when loading the camera we never use pre-dessicated film, but instead load the camera with film that has been at atmosphere (and warmed up to room temperature if it's been in the refrigerator). We then dessicate the camera box prior to putting it in the microscope.
On a related topic, on occasion we've observed a fine network of cracks in the emulsion of our SO163 film. Under an optical microscope the film looks similar to a dried out mud flat with cracks spaced rougly 0.05 mm apart. These cracks would appear regardless of whether we dried the film at room temperature overnight or in a heated film drier. The cracking also appeared with both new and old chemicals (D19 diluted 2:1 4 minutes; kodak rapid fixer 5 minutes). Thinking we had a bad batch of film we sent some of the material to Kodak, but they were unable to duplicate the effect. Finally, it was suggested that we reduce the concentration of hardener in the fixer to half of what Kodak recommends. This seems to have reduced the cracking quite a bit, although not entirely. Has anyone had similar problems?
+---------------------------------------------+ ! Douglas L. Medlin ! ! Physical Properties of Materials Department ! ! Mail Stop 9402 ! ! Sandia National Laboratories ! ! Livermore, California 94551 ! ! ! ! (510) 294-2825 ! ! dlmedli-at-california.sandia.gov ! +---------------------------------------------+ .\
Is there anyone out there who has any tips/techniques on polishing hygroscopic material for microanalysis. Specifically, I need to polish some CaO particles with CaC rinds. I could probably polish them with silicone oil, but how do I get the oil off the samples before I put them in the microprobe? Any comments or suggestions would be appreciated.
Thanks in advance
Glenn ********************************************************************** * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * * Electron Microprobe Lab Phone: (514) 398 6774 * * Earth and Planetary Sciences Fax: (514) 398 4680 * * McGill University THERE ARE THREE SIDES TO EVERY STORY; * * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * **********************************************************************
I am a graduate student currently working on a project utilizing EDS to get quantitative elemental information of atmospheric particles. In past few monthes, I have been trying to get hold of a article cited below through the interlibrary service and without any success. It will be greatly appreciated if anyone out there can direct me where and how to get this article.
Heinrich, K. F. J. (1987) "Mass Absorption Coefficients for Electron Probe Microanalysis." in "Proc. 11th Int. Cong. on X-ray Optics and Microanalysis." edited by J. Brown and R. Packwood, published by J. D. Brown, University of Western, Ontario, pp67-119.
Po-Fu Huang Particle Technology Laboratory Department of Mechanical Engineering University of Minnesota (Office) (612) 626-7227 (Lab) (612) 625-7307 (Fax) (612) 625-6069
We pre-pump negatives in a separate vacuum chamber with phosphorus pentoxide powder placed in the bottom of chamber. This film is loaded into a spare cassette and has always been suitable for use after being pumped for 1-2 hours or left under vacuum. After about a week, the powder absorbs moisture and must be replaced. The old powder can be neutralized with NaOH pellets and when NaOH pellets have dissolved, can be discarded by flushing with water.
Marge Hukee EM Core Facility hukee.margaret-at-mayo.edu Mayo Foundation (507) 284-3148 ---------------------------------------------------------------------------- --
I am currently writing a grant proposal, and I have been knocking my head against the wall looking for some sort of data on the optical transmission properties of nervous tissue (the brain). Does anyone have a good reference, or, from a practical standpoint, how thick do brain tissue slices have to be to obtain good optical transmission (esp. relative to other fatty tissue slices like those from the earlobe, whose transmission characteristics I can find since people have been doing pulse oximetry.).
Thanks in advance,
John Conroy University of Utah Dept. Bioengineering
We had some static discharge on our 35 mm film in the past. It occurred when pre-desiccated bulk was being rolled onto cassettes. The problem disappeared after bulk film was left in atmosphere. We never had problem with plates because we did not pre-desiccate plates. One may try leaving the exposed plates in a humid chamber for a while before developing them, if the problem persisted.
I have an LKB Ultratome III that is surplus to our needs. Purchased in 1978, almost never used. If you might be interested in making an offer or know of a worthwhile place for a donation, please let me know.
Thanks,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95060 emlab-at-ucsco.ucsc.edu (408) 459-2477
Glenn (and others) I have polished B2O3 with the 3M lapping film and no water. It doesn't produce a great polish, but you can usually find a few spots where you can squeeze a beam in. If you don't use the 3M film, let me know and I'll tell you where you can get some or even send you a scrap. Maggy Piranian M.U.N.
On Fri, 24 Feb 1995, Glenn Poirier wrote:
} Greetings Microscopists } } Is there anyone out there who has any tips/techniques on polishing hygroscopic } material for microanalysis. Specifically, I need to polish some CaO } particles with CaC rinds. I could probably polish them with silicone } oil, but how do I get the oil off the samples before I put them in } the microprobe? Any comments or suggestions would be appreciated. } } Thanks in advance } } Glenn } ********************************************************************** } * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * } * Electron Microprobe Lab Phone: (514) 398 6774 * } * Earth and Planetary Sciences Fax: (514) 398 4680 * } * McGill University THERE ARE THREE SIDES TO EVERY STORY; * } * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * } ********************************************************************** }
EM folk: I'm trying to use T-max 100 in the 35mm camera on my Zeiss EM 10C. Everything works well (you get nice short exposures, and this particular camera can be handled in the light, so there's no problem with using a panchromatic film). But the film gets *BRITTLE* in high vacuum. I have snapped the film in half simply by bending it as I unloaded the camera. Any hints, or do I just need to switch to one of the orthochromatic films? If so, which one(s) have you tried and liked? Julian Smith III Biology Winthrop University Rock Hill, SC 803-323-2111 (vox)
Message-ID: {MAILQUEUE-101.950227084552.256-at-physics.uwc.ac.za} To: microscopy-at-aaem.amc.anl.gov
Douglas Medlin wrote about fine cracks appearing on some of their TEM negatives. I believe you are experiencing what photographers called :crazing:. This is an effect that occurs when the temperature of the developer and fixer is not the same, or at least the difference is not small enough. It is exactly as you describe it, a fine line structure on the film that looks that mud cracks.
Dirk Knoesen Department of Physics, University Western Cape, Bellville, 7530 South Africa e-mail: dirk-at-physics.uwc.ac.za Phone: (+21) 959 2236 Fax: (+21) 959 3474
We use Eastman motion picture film (5302 fine grain release positive film) in Philips EM300 in the past and currently in Zeiss EM902 without any problem.
One suggestion I would make would be polishing with abrasive lapping films. These are abrasive particles which are bonded to a polyester film. It is a higher grade of traditional SiC grinding paper and it comes in micronized particle sizes. The material is designed to be used either with or without a lubricant which should take care of your problem.
While it is always preferable to polish with some sort of lubricant, this is a viable alternative for anhydrous materials. I must also add that the lapping film is one of our products so I do have a vested interest in this suggestion. If you would like a sample to try out, please contact me. It is avaialable in Aluminum oxide down to .05 micron and in Diamond down to 0.1 micron. Typical price for Al2O3 film is about $1.39 per 8" PSA Disc and for Diamond about $24 per 8" PSA disc. 8" Plain Back Diamond Film is more typically used and is priced at about $20 per 8" Disc. Prices, of course, will decrease in quantity. The diamond plain back films are used extensively in Tripo Polishing for SEM and TEM applications.
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
I have used Kodak Fine Grain Release Positive (5302) in Philips microscopes for 30 years. Never gave a problem with cracking with the unperforated (no longer available) or perforated stock. Gave some problems with "lightning strikes" on the final 4-5 frames out of 40. Philips have special cameras with large diameter hubs so the film is never bent much and used unperforated film so there was no tendency to crack at perforations. A 35 mm camera in our Hitachi H-7000 gives very fine cracking across the film between the edges of the perforations even when using the film Hitachi recommends. This can only be avoided by using film which is only JUST dry enough, shooting the whole roll and processing it in about 4 hours.
For your information, Materials Microscopy is a new newsletter dedicated to the professional interests of the working materials microscopist. Those who wish to receive a free subscription should provide us with their full mailing address via FAX -at- (602) 947-7615 or E-mail: MatlsMicrs-at-aol.com. Contributed articles should be mailed to: Materials Microscopy P.O. Box 2014 Scottsdale, AZ 85252 Please contact me if you need any further information. Thank you.
Rene E. Nicholas Circulation Manager Materials Microscopy
Hi Barbara! I tried to no avail to send you the present message at your email address: it stubbornly bounced back, so I eventually post it to the microscopy list. I apologize to the members of this list if they should feel this is wasting bandwidth: just delete and go on to the next message ;-)
Barbara! I am afraid I did not make my question clear enough ;-) What I wanted to know was: 1) the testes you wish to fix are from what species ?
2) for what kind of microscopy are they to be processed ? I.e. T.E.M., S.E.M., or light microscopy ?
3) What's the embedding medium?
4) in the case of light microscopy, do you contemplate using some special technique (besides for recognizing the different stages of the seminiferous epithelium) which would require that some specific chemical would be either required or, on the contrary, avoided?
In my personal experience with rat testes and light microscopy (this is rather very long ago: more than 30 years! :-( ), Bouin and other picric acid containing variations thereof are not optimal fixatives for preservation of the acrosomial apparatus of spermatids, which is one of the features used by most classification methods (Clermont's among others...). The best fixatives for light microscopy of paraffin embedded tissue are those containing potassium dichromate such as, for example, Helly's fluid (Zenker + formaldehyde), but they also are followed, after rinsing and dehydration, by shrinkage which is still more important than that observed after Bouin's or Bouin-Hollande's (somewhat better) fluids.
BTW, what's wrong with picric acid? It is always shipped in 'moist condition' in order to minimize hazards. I work in our lab since 1953, with Bouin, Bouin-Allen, Bouin-Hollande, Dubosq-Brazil etc., every day, and we never experienced any problem. We store the picric acid for our needs in the form of a saturated aqueous solution which keeps for as long as you will. This stock solution is then used for preparing all picric acid containing fixatives.
There is no point in trying to eliminate the picric acid from the fixed pieces of tissue before sectioning. If the presence of picric acid in the slides should hamper subsequent staining (a rare occurrence, btw), then the easiest and expeditious way to get rid of it is to pretend your tissue was fixed with a sublimate containing fluid: at the rehydration step, your slides should undergo treatment by alcoholic iodine (or lugol solution) and sodium thiosulfate in the usual way and 'voila': no more picric acid on the slides within 2 minutes! Under no circumstances should the pieces of tissue fixed with a picric acid containing fixative undergo rinsing in water! This is sheer heresy ;-) because it induces a lot of swelling in the tissue before shrinking even more during dehydration (I never found out which histologist ever advocated such method, but I know quite a number of renowned ones who firmly condemned it, and with good reason). If you insist on rinsing the pieces of testes after fixation, then merely increasing the number of steps through 90x alcohol should do the trick; you may even add a thin layer (1-2 mm thick) of lithium carbonate on the bottom of the alcohol containing flasks: this helps dissolving part of the picric acid away.
Possibly the method which should yield the best pictures has been documented in a book entitled 'Histological and histopathological Evaluation of the Testis', by Russell,Ettlin, Sinha Hikim and Clegg: Cache River Press 1990, (15777 Bolesta, Box 129, Clearwater, Fl 34620): ISBN 09627422-0-1.
At the end of the book, there is an appendix about the materials and methods which you might find useful (although perfusion fixation of rat testes is among the most frustrating experiences I ever had, even with a lot of heparin ). This would also require embedding in epoxy resin and making large semi-thin sections, which I don't know whether you are prepared to do ;). Glutaraldehyde certainly seems to me the best choice, but then paraffin embedding would be ruled out (too hard and too brittle after paraffin embedding). HTH, and good luck. Let me know about the outcome (good, I hope) John (correction: alcohol above should read 90 percent in volume)
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
To whomever, Would someone please repost the recent info on lipid sizing (K.Walters?) and the three responses on lipid negative staining. I had another crash and lost these responses. Thanks Mike D.
I am currently writing a grant proposal, and I have been knocking my head against the wall looking for some sort of data on the optical transmission properties of nervous tissue (the brain). Does anyone have a good reference, or, from a practical standpoint, how thick do brain tissue slices have to be to obtain good optical transmission (esp. relative to other fatty tissue slices like those from the earlobe, whose transmission characteristics I can find since people have been doing pulse oximetry.).
Thanks in advance,
John Conroy University of Utah Dept. Bioengineering
Dear John,
It has been years since I worked in this area, but I suggest you might look at the literature for the optical transmission properties of retina. As a nervous tissue, its optical properties should be quite similar to the CNS as long as the measurements are not made in the vicinity of the fovea. And, of course, due the importance of retinal transparency for vision, its optical properties must be well known. For a place to start I'd have a look at the "Handbook of sensory physiology" Springer-Verlag.
Just my two cents worth, Steven ----------------------------------------------------------------------------- Steven L. Goodman, Ph.D. Dept. Animal Health and Biomedical Sciences 608-262-0816 (office) University of Wisconsin 608-262-7420 (FAX) 1655 Linden Drive Madison, WI 53706 SLG-at-AHABS.WISC.EDU --------------------------------------------------------------------------
} Return-Path: {delannoy-at-welchlink.welch.jhu.edu} } Received: from AAEM.AMC.ANL.GOV by welchlink.welch.jhu.edu (5.0/SMI-SVR4) } id AA04845; Tue, 28 Feb 1995 09:52:41 -0500 } Date: Tue, 28 Feb 1995 09:38:53 -0500 } Message-Id: {9502281438.AA02618-at-welchlink.welch.jhu.edu} } X-Sender: delannoy-at-welchlink.welch.jhu.edu } Mime-Version: 1.0 } To: microscopy-at-aaem.amc.anl.gov } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } Subject: lipid sizing and neg. stain } X-Mailer: {Windows Eudora Version 1.4.2b16} } Content-Type: text/plain; charset="us-ascii" } } To whomever, } Would someone please repost the recent info on lipid sizing (K.Walters?) } and the three responses on lipid negative staining. I had another crash and } lost } these responses. } Thanks } Mike D. } }
Just a note to let you guys know that our link from this mailing list to the Usenet newsgroup sci.techniques.microscopy has been broken. The gateway at sci.techniques.microscopy.usenet-at-decwrl.dec.com has been closed down. Dec no longer support it. If you want as much coverage for you questions and posts as possible I encourage you to post both to this list and also to the Newssgroup.
If anyone knows how to set up a mail gateway to the Newsgroup, and also from the Newsgroup to the mailing list, I would be very interested to hear how to do it so we could reestablish the link and possibly make it bi-directional this time!
Many thanks Jfm.
______________ John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
I tried to no avail to send you the present message at your email address: it stubbornly bounced back, so I eventually post it to the microscopy list. I apologize to the members of this list if they should feel this is wasting bandwidth: just delete and go on to the next message ;-)
Barbara! I am afraid I did not make my question clear enough ;-) What I wanted to know was: 1) the testes you wish to fix are from what species ?
2) for what kind of microscopy are they to be processed ? I.e. T.E.M., S.E.M., or light microscopy ?
3) What's the embedding medium?
4) in the case of light microscopy, do you contemplate using some special technique (besides for recognizing the different stages of the seminiferous epithelium) which would require that some specific chemical would be either required or, on the contrary, avoided?
In my personal experience with rat testes and light microscopy (this is rather very long ago: more than 30 years! :-( ), Bouin and other picric acid containing variations thereof are not optimal fixatives for preservation of the acrosomial apparatus of spermatids, which is one of the features used by most classification methods (Clermont's among others...). The best fixatives for light microscopy of paraffin embedded tissue are those containing potassium dichromate such as, for example, Helly's fluid (Zenker + formaldehyde), but they also are followed, after rinsing and dehydration, by shrinkage which is still more important than that observed after Bouin's or Bouin-Hollande's (somewhat better) fluids.
BTW, what's wrong with picric acid? It is always shipped in 'moist condition' in order to minimize hazards. I work in our lab since 1953, with Bouin, Bouin-Allen, Bouin-Hollande, Dubosq-Brazil etc., every day, and we never experienced any problem. We store the picric acid for our needs in the form of a saturated aqueous solution which keeps for as long as you will. This stock solution is then used for preparing all picric acid containing fixatives.
There is no point in trying to eliminate the picric acid from the fixed pieces of tissue before sectioning. If the presence of picric acid in the slides should hamper subsequent staining (a rare occurrence, btw), then the easiest and expeditious way to get rid of it is to pretend your tissue was fixed with a sublimate containing fluid: at the rehydration step, your slides should undergo treatment by alcoholic iodine (or lugol solution) and sodium thiosulfate in the usual way and 'voila': no more picric acid on the slides within 2 minutes! Under no circumstances should the pieces of tissue fixed with a picric acid containing fixative undergo rinsing in water! This is sheer heresy ;-) because it induces a lot of swelling in the tissue before shrinking even more during dehydration (I never found out which histologist ever advocated such method, but I know quite a number of renowned ones who firmly condemned it, and with good reason). If you insist on rinsing the pieces of testes after fixation, then merely increasing the number of steps through 90x alcohol should do the trick; you may even add a thin layer (1-2 mm thick) of lithium carbonate on the bottom of the alcohol containing flasks: this helps dissolving part of the picric acid away.
Possibly the method which should yield the best pictures has been documented in a book entitled 'Histological and histopathological Evaluation of the Testis', by Russell,Ettlin, Sinha Hikim and Clegg: Cache River Press 1990, (15777 Bolesta, Box 129, Clearwater, Fl 34620): ISBN 09627422-0-1.
At the end of the book, there is an appendix about the materials and methods which you might find useful (although perfusion fixation of rat testes is among the most frustrating experiences I ever had, even with a lot of heparin ). This would also require embedding in epoxy resin and making large semi-thin sections, which I don't know whether you are prepared to do ;). Glutaraldehyde certainly seems to me the best choice, but then paraffin embedding would be ruled out (too hard and too brittle after paraffin embedding). HTH, and good luck. Let me know about the outcome (good, I hope) John (correction for above glitch about the alcohol/ethanol: should read alcohol 90 percent (in volume); I am still stuck with kermit and an awfull editor :( ).
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
this just is a test because it seems I am not able to reach the list. Please ignore and delete. I apologize for the inconvenience. desclinj-at-ulb.ac.be
The 1 MeV AEI EM-7 at the University of Wisconsin, Madison has been used for the study of biological specimens for the past 23 years. In the early 1980's, almost every subsystem of this instrument (magnetic shielding and ambient field, lens-current and HV stability, vibration isolation, LaB6 source, TV-interface, Axis-centered stereo-tilt stage, high-resolution cold-stage etc.) was upgraded with a view to performing high-resolution studies on frozen biological specimens. Images of oriented gold films at that time showed 0.14 nm spots in bright field and using an 11 mm gap (Pawley, J.B. (1984) Ultramicroscopy 13-4:387-406, describes most of these improvements in detail.)
Because it seems probable that the present management of this instrument will decide (has decided?) to decommission it in the very near future, (It was almost scrapped over last Christmas), I am posting this notice in case there are those that feel that the country has need of a second instrument with the general specifications of the Berkeley ARM or even of the very-stable Haefely 1-MEV supply that it contains.
The instrument is in operating condition, but, until recently, has received less maintenance than it should have for the past five years or so.
The questions are:
1. Do you have important projects that require higher resolution (or a large gap) than that available from 200-400kV instruments? (Keep in mind that knock-on damage may be more severe at the higher voltage.) 2. Would you be willing to travel to Madison to obtain such facilities? 3. Would you be interested in relocating the instrument to a more convenient location?
Bear in mind that, because the instrument requires a room 3 floors high and a large (100 t) vibration-isolation block and also has substantial power and cooling requirements, moving it would be a bit complex. However, it could surely be done at less than 10% of $5M cost of the new Stuttgart 1.25 MeV instrument.
It would also need stage-rods more suited for material-science applications but it is possible that the stage formerly fitted to the Cambridge HREM could be fitted without great trouble as both instruments used the same column.
The purpose of this communication is solely for the information of possible users with the aim of avoiding anyone in the future saying "If only you had told me?". I do not represent the Integrated Microscopy Resource where the instrument is installed.
Please send any responses directly to me and do so ASAP:
jpawley-at-macc.wisc.edu (James Pawley)
***************NEW ADDRESS************** Prof. James B Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr. Madison, Wisconsin, 53706. JPAWLEY-at-MACC.WISC.EDU
Dr. Keith Moulding, Materials Characterisation and Preparation Centre, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 43-22 85 61 89 (work) + 43-22 43 83 23 (privat) Fax.: + 43-22 85 47 26 e-mail.: abrech-at-bio.uio.no
Please, I would like to join the list. Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 43-22 85 61 89 (work) + 43-22 43 83 23 (privat) Fax.: + 43-22 85 47 26 e-mail.: abrech-at-bio.uio.no
Does anyone have a referrence for backscattering cross-sections of electrons in various substances as a function of energy and/or of angle? I'd really appreciate a reply either to the list, by email or if I could get a table by fax at (518) 474-8590. TIA. Yours, Bill Tivol tivol-at-tethys.ph.albany.edu
I'm looking for any recomendations for freeze-fracture units. We're putting together a grant here to obtain one and the only souce I've come up with is Bal-tec. I am looking for other possible vendors. Apparently JEOL has stopped production of their freeze- fracture unit. Specifically we're looking for units comparable to the Bal-Tec BAF 060 (not that I as of yet have anything against the BAF 060, but it is nice to look around first.)
Thank you.
Richard E. Edelmann Electron Microscopy Facility Supervisor Miami University, Oxford, Ohio
I don't think there is a unit out as yet that can match the specs and design of the new BAL-TEC unit. Cressington is another company which manufactures freeze-fracture unit. While it may not have all the features of the new BAL-TEC one, it is reported to be good for routine freeze-fracture.
************************************************************************* M.V. Parthasarathy Professor of Plant Biology Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14850 Telephone: (607) 255-1734 Fax: (607) 255-5407 E-mail: mvp2-at-cornell.edu ***********************************************************************
} I'm looking for any recomendations for freeze-fracture units. } We're putting together a grant here to obtain one and the only souce } I've come up with is Bal-tec. I am looking for other possible } vendors. Apparently JEOL has stopped production of their freeze- } fracture unit. Specifically we're looking for units comparable to } the Bal-Tec BAF 060 (not that I as of yet have anything against the } BAF 060, but it is nice to look around first.) } } Thank you. } } Richard E. Edelmann } Electron Microscopy Facility Supervisor } Miami University, Oxford, Ohio
It looks as though you have some misinformation. JEOL is making the JFD 9000 series freeze-fracture instruments. I know that John Rash has recently purchased and installed the 9010CR with the latest modifications, including a specimen stage with rapid cooling.
Both Bal-Tec and JEOL make fine instruments and are worth looking at.
John chandler-at-lamar.ColoState.EDU Fort Collins CO
Many published investigations on stress voids in aluminum/silicon alloys report using CF4/O2 plasma as the deprocessing method to remove the final PECVD oxynitride/ SiN. Does the use of this plasma chemistry generate its own voids by removing silicon precipitates which in turn leave voids possibly identified as stress voids? Precipitates that were in or near the correct orientation give the appearance (typically wedge-shaped) of a stress void. Optical inspection without deprocessing does not have adequate resolution. I'd appreciate any comments on the CF4/O2 method and a possible alternative method (other than argon backsputtering).
One of the divisions of our lab teases apart sural nerve tissue that have been kept in a 1:1 mixture of epon:propylene oxide. The samples are very easy to tease when the mixture is very fresh. The problem is that samples became back-logged and were not able to be teased right away. A quick decision had to be made, and the samples were frozen at -70C until they could be teased.
The nerve fibers need to be infiltrated to be teased but not polymerized or else they become too fragile. A problem is becoming evident that after a long period of storage, the nerve samples are being fully infiltrated and polymerized despite being frozen (which we thought would almost stop the polymerization process because it is largely dependant on heat). Realizing that hind-sight is 20/20, would it have been better to leave the accelerator out of the epoxy mixture? What affect would this have on the quality of the resin? I understand accelerator to act as a catalyst being a part of the polymerization reaction but not really consumed. Any opinions to offer?
I have been asked to make a brief (15 min) presentation at Scanning 95 in Monterey, on March 30, 1995. The topic is 'Changing Role of the Microscopy Lab in the University'. I know how my lab has changed, but would like to get some idea from a broader sample of microscopists about changes in our role etc.
My rough observations at this time include some of the following. More labs seem to be consolidating, so we keep trying to catch up with too many techniques, especially as staffs are reduced.
There are lots of other places for researchers to spend their funding than before, we have lost lots of users in biology to molecular techniques. Some of the new faculty see microscopy as 'too hard'.
There are fewer and fewer students who want to be 'microscopists'. Most seem to want to come in on Monday, drop off something, and return on Friday with a picture or two for their thesis.
The EM lab on our campus has evolved into an imaging and microscopy lab. Because of our interest in images, we have always had the best darkrooms, now we are trying to lead the way into digital imaging. This means more computers and branching out from traditional 'microscopy'.
I could probably think of other changes, but that wouldn't leave anything for you to add. Don't be shy, reply directly to me and I will try to incorporate your comments in my presentation in a constructive way. Let me know if you want specific credit for an idea, or if you prefer to have your input blended for anonymity.
If you can't make it to Monterey, I'll send a summary if you ask.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95060 emlab-at-ucsco.ucsc.edu (408) 459-2477
Reply to: RE} Re: Optical Microscopy of Tissues Dear John, Sorry about the delay in responding - I missed your message in the deluge I received while visiting labs for a few days. Check out the following papers on infrared DIC-videomicroscopy.
H.-U. Dodt, W. Zieglgansberger (1990) Brain Research 537:333-356; (1994) Trends in Neurosciences 17 (11): 453-458 (and references therein).
They looked with transmitted light at depths of up to 100 um in 300 um thick sections. Used a 63x 1.4 NA oil immersion objective and DIC optics to visualize nerve terminals. Note that the Nikon 60x 1.2 NA WATER immersion objective is now available and has advantages in deep sections (see for example Mel Brenner's application note in the November 1994 Journal of NIH Research).
Infrared penetrates well because water and tissues doesn't absorb it as much as visible. For video work a CCD (or cooled CCD) is used because they are sensitive to IR (unlike most tube cameras and your eye). DIC is the method of choice because of optical sectioning. A general review or IR cameras is: Silverman et al (March 1992) Scientific American 78-83 and 112-113. Any high quality video CCD camera will work (as long as it does not have an IR blocking filter!).
Sincerely,
Dr. George McNamara Universal Imaging Corporation --------------------------------------
Hello,
I am currently writing a grant proposal, and I have been knocking my head against the wall looking for some sort of data on the optical transmission properties of nervous tissue (the brain). Does anyone have a good reference, or, from a practical standpoint, how thick do brain tissue slices have to be to obtain good optical transmission (esp. relative to other fatty tissue slices like those from the earlobe, whose transmission characteristics I can find since people have been doing pulse oximetry.).
Thanks in advance,
John Conroy University of Utah Dept. Bioengineering
--------- Dear John,
It has been years since I worked in this area, but I suggest you might look at the literature for the optical transmission properties of retina. As a nervous tissue, its optical properties should be quite similar to the CNS as long as the measurements are not made in the vicinity of the fovea. And, of course, due the importance of retinal transparency for vision, its optical properties must be well known. For a place to start I'd have a look at the "Handbook of sensory physiology" Springer-Verlag.
The objective lens and the upper and lower column parts of our 20 year old side entry JEOL 100 C microscope need replacement. New parts, however, are rather expensive in view of the age of the instrument so we're looking for anyone who has some second hand spare parts for sale. Sincerely, Nick Schryvers tel: 32-3-2180247 fax: 32-3-2180257 EMAT, University of Antwerp Groenenborgerlaan 171 B-2020 ANTWERP Belgium
I first became aware of the recomendation to replace the epoxy resin= acelerator DMP-30 with the acelerator BDMA when Audrey Glauert published= her article "Accelerators for epoxy resins" in the RMS Proceedings (Sept, 1= 987).
The reason offered to encourage the switch from DMP-30 to BDMA was the fact= that BDMA is less viscous, therefore diffuses into the tissue sample= better. This gives a more even polymerisation and better sectioning. BDMA= also has a longer shelf life.
I followed Glauert's recomendation, replacing DMP-30 with BDMA in our resin= formula's. However, Audrey Glauert did not mention in the article that= we should double the quantity of BDMA in the formulation when switching= from DMP-30 to BDMA.
I replaced the DMP-30 with BDMA, using the BDMA at the same concentration as= I had used the DMP-30.
Generally, it worked fine. Occasionally I had 'strange embedding' problems,= ie samples not properly polymerised.
It then came to my attention that I should be using BDMA at twice the DMP-30= concentration. I did not pay much attention to this at the time as= generally everything had been working fine and I was to busy to experiment= with the change.
After recent 'flow' on the List server the topic again surfaced i.e. I= should be using BDMA at twice the DMP-30 concentration. This I have now tr= ied.
My problem is that if I do an overnight infiltration in resin with the= increased concentration of BDMA the resin is so viscous next morning it= won't even come out of the vial when tipped upside down. To change to= fresh resin is almost impossible, in fact it is easier to change the= samples to fresh resin in a new processing tube. Unfortunately even then= it is not totally successful as a large 'blob' of tacky resin stays with= the sample.
I did a little experiment as outlined below;
1. Made up resin with normal DMP-30 concentration. 2. Made up resin with BDMA at normal DMP-30 concentration. 3. Made up resin with BDMA at twice DMP-30 concentration.
Results
Viscosity (by eye) Colour To start with DMP-30 most viscous slightly orange BDMA (-at- DMP-30 conc) least viscous straw coloured BDMA (-at- 2 x DMP-30 conc)similar to BDMA -at- DMP 30 conc straw coloured
After 5 hours DMP-30 least viscous darker straw coloured BDMA (-at- DMP-30 conc) similar to DMP-30 straw coloured BDMA (-at- 2 x DMP-30 conc)quite viscous straw coloured
Overnight DMP-30 least viscous straw coloured BSMA (-at- DMP-30 conc) slightly more viscous than DMP-30 straw coloured BDMA (-at- 2 x DMP-30 conc)very very viscous straw coloured
After the overnight step both the DMP-30 mix and the BDMA mix at the DMP-30= concentration were quite acceptable, ie no difficulty replacing the old= resin with fresh resin.
The BDMA mix at the twice DMP-30 concentration ws very very tacky.
After polymerisation both the DMP-30 and the BDMA -at- the DMP-30= concentration are a light straw colour (normal expectation). The BDMA -at-= twice the DMP-30 concentration was slightly darker, although still an= acceptable straw colour.
By modifying my processing schedule such that the samples stay in 3 parts= plastic and 1 part propylene oxide overnight I can successfully process a= sample however, I am a little unhappy with the short time the sample is in= 'resin only' the following day, ie out of 3;1 and into fresh plastic first= thing in the morning, embedded and in the oven before I go home. With a= lot of samples to embed at times trying to give the samples as long as= possible in the 'resin only' makes the end of the day a bit tight for time= .
The problem becoming particularly accute when using our Lynx Automatic= Tissue processor. If using BDMA -at- twice the DMP-30 concentration I cannot= leave the resin in the processer overnight as it is to tacky the next day.
My Question ??? (at last some may say)
1. How are others out there, who are using BDMA at twice the DMP-30= concentration getting around the problem of the resin going very tacky= overnight, during infiltration?
What sort of infiltration times do you use?
2. Are any of you using BDMA at twice the DMP-30 concentration in an= automatic tissue processor?
3. Is any one else having problems with the resin if made up with BDMA at= twice the DMP-30 concentration?
Thank you in anticipation.
Allan Mitchell South Campus Electron Microscope Unit Department of Anatomy and Structural Biology Otago Medical School
Dennis Shubitowski {dennis-at-odin.morph.med.umich.edu}
Dennis,
If you omit the accelerator, it would probably work well. We use Epon-Araldite (Polyscience kit), and we routinely store the mixed resin (Epon + Araldite + DDSA), without catalyst (DMP-30), in a 4 degree C refrigerator. It will remain good for at least 6 months. When we are ready to embed, we measure out whatever volume we want, add DMP-30 to make 2%, mix, and it is fine. If you stored your nerve samples in 1:1 Epon (no accelerator):PO, and later decided to embed them, you could then embed them by standard procedures with catalyzed resin. If you have any questions about this, give me a call (763-1287), or drop over (2 blocks away).
Kent
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School Ann Arbor, MI 48109-0616 {akc-at-umich.edu}
----------------------------------------
On Thu, 2 Mar 1995, Dennis Shubitowski wrote:
} One of the divisions of our lab teases apart sural nerve tissue that have } been kept in a 1:1 mixture of epon:propylene oxide. The samples are very } easy to tease when the mixture is very fresh. The problem is that samples } became back-logged and were not able to be teased right away. A quick } decision had to be made, and the samples were frozen at -70C until they } could be teased. } } The nerve fibers need to be infiltrated to be teased but not polymerized } or else they become too fragile. A problem is becoming evident that after a } long period of storage, the nerve samples are being fully infiltrated } and polymerized despite being frozen (which we thought would almost stop } the polymerization process because it is largely dependant on heat). } Realizing that hind-sight is 20/20, would it have been better to leave the } accelerator out of the epoxy mixture? What affect would this have on the } quality of the resin? I understand accelerator to act as a catalyst being } a part of the polymerization reaction but not really consumed. Any } opinions to offer? } } Thanks, } } Dennis } } } } }
MICROSCOPY-at-AAEM.AMC.ANL.GOV Cc: Dennis Shubitowski {dennis-at-odin.morph.med.umich.edu} Message-id: {01HNOZPUD02Q93M845-at-gnv.ifas.ufl.edu} MIME-version: 1.0 X-Mailer: Windows Eudora Version 1.4.4 Content-type: text/plain; charset="us-ascii" Content-transfer-encoding: 7BIT
In reference to Dennis Subitowksi's question about expoxt accelerator, I can say that we routinely exclude the accelerator during infiltration with Epon, Epon/Araldite mixture and Epon substitutes and add it only to the second pure resin infiltration step. This allows long infiltrations in earleir steps with difficult specimens. When we have compared adding it to all steps, the results seem to be the same. Hope that helps
*****ORIGNINAL MESSAGE BELOW******** } One of the divisions of our lab teases apart sural nerve tissue that have } been kept in a 1:1 mixture of epon:propylene oxide. The samples are very } easy to tease when the mixture is very fresh. The problem is that samples } became back-logged and were not able to be teased right away. A quick } decision had to be made, and the samples were frozen at -70C until they } could be teased. } } The nerve fibers need to be infiltrated to be teased but not polymerized } or else they become too fragile. A problem is becoming evident that after a } long period of storage, the nerve samples are being fully infiltrated } and polymerized despite being frozen (which we thought would almost stop } the polymerization process because it is largely dependant on heat). } Realizing that hind-sight is 20/20, would it have been better to leave the } accelerator out of the epoxy mixture? What affect would this have on the } quality of the resin? I understand accelerator to act as a catalyst being } a part of the polymerization reaction but not really consumed. Any } opinions to offer? } } Thanks, } } Dennis } } } } } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
Due to renovations within the department, and the purchase of a confocal microscope, we have 2 transmission EMs in excess of our needs that we would like to move. Both microscopes are in good working condition, one has been under a service contract until this past year (though lightly used). The scopes:
1. Hitachi H-600 STEM (the one under service contract until recently)
2. Zeiss EM-9S TEM
We are not quite at a "No reasonable offer refused!" price level, but we would like to see these scopes get some more use and we're eager to make use of the space they are now occupying. So....if you are at all interested, contact me and we can discuss the particulars and terms of sale.
TIA,
Phil Rutledge voice: (410) 455-3582 Email: prutle1-at-gl.umbc.edu Fax: (410) 455-3875
Dennis Shubitowski {dennis-at-odin.morph.med.umich.edu}
We had a similar experience some years ago. Not with nerve, but with other material. I can offer the following humble comments.
1. As you learned, resin does polymerize in even at freezing temperatures. We have not tried -70 C and you did not specify how far below freezing you stored your specimens. We did not test exactly why, but reasoned that since the reaction is highly exothermic and since both resin and tissue are good insulators, it is probable that local areas are heated enough to allow polymerization. I look forward to any comments on the errors in our reasoning, since this was only a guess to explain the results. 2. Leaving out the accelerator does tremendously slow down the reaction, but polymerization still occurs in absence of accelerator, and even in the cold, so don't store for grossly extended times. To polymerize infiltrate tissue with fresh resin containing accelerator (we use 2 changes, 4 hours each) after thawing tissue. This works well, although some areas may have obvious interfaces between resins of different hardness.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Thu, 2 Mar 1995, Dennis Shubitowski wrote:
} One of the divisions of our lab teases apart sural nerve tissue that have } been kept in a 1:1 mixture of epon:propylene oxide. The samples are very } easy to tease when the mixture is very fresh. The problem is that samples } became back-logged and were not able to be teased right away. A quick } decision had to be made, and the samples were frozen at -70C until they } could be teased. } } The nerve fibers need to be infiltrated to be teased but not polymerized } or else they become too fragile. A problem is becoming evident that after a } long period of storage, the nerve samples are being fully infiltrated } and polymerized despite being frozen (which we thought would almost stop } the polymerization process because it is largely dependant on heat). } Realizing that hind-sight is 20/20, would it have been better to leave the } accelerator out of the epoxy mixture? What affect would this have on the } quality of the resin? I understand accelerator to act as a catalyst being } a part of the polymerization reaction but not really consumed. Any } opinions to offer? } } Thanks, } } Dennis } } } } }
Richard- Why not try infiltrating your samples overnight in 100% resin with the lower concentration of BDMA, then swithching to a "short" infiltration with fresh resin made with 2X concentration BDMA. then embedding in the higher concentration of BDMA in fresh resin? just a hunch. -Mike
I performed a similar set of experiments: BDMA vs DMP-30. Although it infiltrated specimens adequately and although they sectioned fine; dealing with changes of 100% plastic that were sooooo viscous was more than I could tolerate. In spite of it's initial higher viscosity and shorter shelf life, I switched back to using DMP-30 in our PolyBed 812 resin. Doing changes of 100% plastic is no longer a headache and something to be dreaded. If someone has a solution to the overnight BDMA-plastic blob problem, I'd like to hear as well.
Cheers...........JohnA
___________________________________________________ | | | John G. Aghajanian, Ph.D. | | Worcester Foundation for Experimental Biology | | 222 Maple Avenue | | Shrewsbury, MA 01545-2737 | | | | Tel: 508 842-8921 ext. 147, 161 | | Fax: 598 842-9632 | | JOHNA-at-sci.wfeb.edu | | | |_________________________________________________|
This message came to me and I have no good answers. Any comments will be appreciated and I will forwrad to the questioners = =Greg, = =Tom Mareci suggested that you might be able to give us some technical =advice on EM's to help us solve a temporary problem we're having. = =We're wanting to move our 4.7T magnet to a location in the ARB basement =120' from Dr. Craig Tisher's EM facility, which has a Zeiss EM 10 SEM =and a Topcon/International Scientific Instruments DS-130C TEM. The SEM =specs {= 3 mG AC and {= 5 mG DC. The TEM specs {=6 mG AC. Dr. Tisher =is reluctant to have us install our magnet, which with shileding is =calculated to produce a DC magnetic field of about 17 mG in the EM room. =Tom and I are having a hard time understanding the DC spec, in light of =the earth's magnetic field being 100x greater than the 5 mG spec. =We are also finding it difficult to get good scientifically sound =answers from the EM manufacturers (at least that we NMR people can =understand!). Could you enlighten us? = =Our main questions are: Will our DC field of 17 mG really pose a problem =for the EM's? If so, how can it be remedied? How much will it cost? =How are these systems currently shielded/compensated for the earth's =magnetic field? (My guess is that at most, a quick realignment of a =passive shield will be all that's required, if there's an observable =effect.) = =Secondarily, since before and after image quality may be the best bottom- =line test of the effect (if any) of our magnet, what standard samples can =you recommend for these Q.C. checks? = =Thanks for your advice and comments. = =-Richard Briggs = =tel: 395-0680 ext 54279 (55-4279) =FAX: 395-0279 =pager: Shands # 3497 =e-mail: rbriggs-at-ufnmr.health.ufl.edu = =P.S.: You may not remember, but I met you at Tom's Christmas party. = ****************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-846-0251 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * ******************************************************************
Hello All, I have a user that is attempting double-label fluroescence for protein localization in sheep ovaries. The ovaries seem to auto-fluoresce very strongly in the FITC channel with some signal in the Rhodamine as well. They have talked to other researchers who have not seen this problem. The tissue is fixed in Carnoy's and paraffin embedded. Has anyone seen or heard of this particular tissue lighting up like a Christmas Tree? TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
In response to my invitation for a free subscription to Materials Microscopy (posted on Feb. 24), we have been receiving a large number of requests and inquiries on a daily basis via e-mail and FAX. We will try to process all your subscription requests prior to the circulation of our next issue. Due to the high volume of mail we have received, I cannot respond to your inquiries individually. However, I have tried to summarize, itemize, and respond to your most frequently asked questions as follows: a. Materials Microscopy is published and circulated at no cost among the materials microscopy community members by Promotech Associates, Inc. b. Our publication is mainly concerned with providing articles and material of interest to working materials microscopist. We would appreciate your contribution which meets this requirement. Further information in this regard can be obtained by contacting our technical editor, Dr. Patricia Labun (a materials microscopist). c. I will send a copy of our "rate sheet" to those of you who expressed interest in placing ads or promotional material. Please feel free to contact me if you require any additional info. Thank you all for your interest in Materials Microscopy.
Rene E. Nicholas Circulation/Sales Manager
Materials Microscopy P.O. Box 2014 Scottsdale, AZ 85252
TEL (602) 947-7603 FAX (602) 947-7615 MatlsMicrs-at-aol.com
I'm wondering if the possible advantages of BDMA (lower viscosity, longer shelf life) are worth all this. I'm still using a 1989 Polysciences Epon-Araldite kit (with DMP-30) that still gives excellent results with standard procedures.
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School {akc-at-umich.edu}
Hi Barbara! I tried to no avail to send you the present message at your email address: it stubbornly bounced back, so I eventually post it to the microscopy list. I apologize to the members of this list if they should feel this is wasting bandwidth: just delete and go on to the next message ;-)
Barbara! I am afraid I did not make my question clear enough ;-) What I wanted to know was: 1) the testes you wish to fix are from what species ?
2) for what kind of microscopy are they to be processed ? I.e. T.E.M., S.E.M., or light microscopy ?
3) What's the embedding medium?
4) in the case of light microscopy, do you contemplate using some special technique (besides for recognizing the different stages of the seminiferous epithelium) which would require that some specific chemical would be either required or, on the contrary, avoided?
In my personal experience with rat testes and light microscopy (this is rather very long ago: more than 30 years! :-( ), Bouin and other picric acid containing variations thereof are not optimal fixatives for preservation of the acrosomial apparatus of spermatids, which is one of the features used by most classification methods (Clermont's among others...). The best fixatives for light microscopy of paraffin embedded tissue are those containing potassium dichromate such as, for example, Helly's fluid (Zenker + formaldehyde), but they also are followed, after rinsing and dehydration, by shrinkage which is still more important than that observed after Bouin's or Bouin-Hollande's (somewhat better) fluids.
BTW, what's wrong with picric acid? It is always shipped in 'moist condition' in order to minimize hazards. I work in our lab since 1953, with Bouin, Bouin-Allen, Bouin-Hollande, Dubosq-Brazil etc., every day, and we never experienced any problem. We store the picric acid for our needs in the form of a saturated aqueous solution which keeps for as long as you will. This stock solution is then used for preparing all picric acid containing fixatives.
There is no point in trying to eliminate the picric acid from the fixed pieces of tissue before sectioning. If the presence of picric acid in the slides should hamper subsequent staining (a rare occurrence, btw), then the easiest and expeditious way to get rid of it is to pretend your tissue was fixed with a sublimate containing fluid: at the rehydration step, your slides should undergo treatment by alcoholic iodine (or lugol solution) and sodium thiosulfate in the usual way and 'voila': no more picric acid on the slides within 2 minutes! Under no circumstances should the pieces of tissue fixed with a picric acid containing fixative undergo rinsing in water! This is sheer heresy ;-) because it induces a lot of swelling in the tissue before shrinking even more during dehydration (I never found out which histologist ever advocated such method, but I know quite a number of renowned ones who firmly condemned it, and with good reason). If you insist on rinsing the pieces of testes after fixation, then merely increasing the number of steps through 90pc x alcohol should do the trick; you may even add a thin layer (1-2 mm thick) of lithium carbonate on the bottom of the alcohol containing flasks: this helps dissolving part of the picric acid away.
Possibly the method which should yield the best pictures has been documented in a book entitled 'Histological and histopathological Evaluation of the Testis', by Russell,Ettlin, Sinha Hikim and Clegg: Cache River Press 1990, (15777 Bolesta, Box 129, Clearwater, Fl 34620): ISBN 09627422-0-1.
At the end of the book, there is an appendix about the materials and methods which you might find useful (although perfusion fixation of rat testes is among the most frustrating experiences I ever had, even with a lot of heparin ). This would also require embedding in epoxy resin and making large semi-thin sections, which I don't know whether you are prepared to do ;). Glutaraldehyde certainly seems to me the best choice, but then paraffin embedding would be ruled out (too hard and too brittle after paraffin embedding). HTH, and good luck. Let me know about the outcome (good, I hope) John
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
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Cressington makes an excellent freeze fracture machine. Suitable for routine work and high resolution shadowing at very low temperatures. In particular the reproducibility, even of W/Ta, is very good, and attainable for everyone (user friendly).
The address is: Dr Peter A Walley CRESSINGTON Scientific Instruments Ltd 24 Chalk Hill Watford Herts WD1 4BX UK Tel 0923 220499 Fax 0923 816646
Success
Marcel Paques Unilever Research Laboratory Vlaardingen The Netherlands E-mail: Marcel.Paques-at-2488PAS.URLNL.Sprint.Com
We use a Cressington CFE- 50B which has proved excellent for reproducible rotary low-angle shadowing of peptides. The system is also very flexible for both W/Ta and Pt/C operation and is easily re-configured for a variety of research applications.
Kevin Jennings SmithKline Beecham Pharmaceuticals UK Microscopy & Flow Cytometry Section The Frythe Welwyn Hertfordshire AL6 9AR United kingdom
I am in the info gathering stage of purschasing a tissue processor for EM samples. I have info on the Reichert-Lynx unit and the one made by RMC. How do you like these units? Comments on advantages and disadvantages are welcome. I also have brouchers on a unit made by Sakura Finetek USA, Inc.. The phone number I called is no longer in working order.(213-539-5441) Is this company still around? These brouchers are probably from the early 80's. Are the any other companys that made tissue processors?
Thanks in advance
Ed Calomeni Medical College of Ohio Toledo, OH
emlab-at-opus.mco.edu
PS Sorry about the post on last friday with no message in the body. Computers can be such good friends.
I recently read W. L. Steffens comments regarding the use of Leighton tubes for embedding cell cultures. Has anyone embedded the coverslips from these tubes in LRWhite resin? I've been having trouble getting consistent polymerization of monolayers on other coverslips (eg Thermanox) and am in search of a new method for obtaining "en face" sections of monolayers for immunocytochemistry. TIA for any hints on the topic.
Christine Powers Dept. of Cell Biology University of Massachusetts Medical Center Worcester, MA 01655
Ed, I can comment on the Lynx tissue processor which we have been using since it first came out, about 8 years ago. The Lynx is made in Australia, and was originally distributed in this country by Fairleigh- Dickinson Laboratories until the distributorship was snatched up by Reichert (now Leica). We paid about 5.5K for the unit and have had little trouble with it aside from a blown power supply (they have upgraded this) and a defective exhaust fan (we fixed this). We use it routinely for processing TEM (epon and L.R. White) and SEM (through dehydration). It works very well is quite consistent. My only complaint involves the expendables for it...vials, caps, baskets, etc. Originally, it was economical to use them once and dispose of them as is intended. Once Leica became the distributor, the price of the expendable essentially tripled...some things quadrupled. As a result, we clean and recycle these components for as long as we can. Additionally, the quality control of these items has suffered immensely. There are now occasional vials that are distorted and won't fit the turntable, and lids and baskets with burrs that must be filed. If you can accept this, I still think that for the money (nearly 9K by now?) its the best one out there. Good Luck.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
We have a JEOL 100CX scanning transmission electron microscope that we no= longer need because of more recent acquisitions. It was maintained continuously= during operation here and was in perfect working condition before we turned it o= ff in =
July, 1994. =
This is a conventional scanning transmission electron microscope with =
resolutions of 3.4 =81 TEM.
It has specimen holders for heating-cooling, double tilt (tilt range is =B1= 45=A1), =
or rotation. =
We would be deligted to entertain offers for it.
Please contact Beth Trend if you are interested.
______________________________________________***************************= ******* Beth Trend trend-at-cems.umn.edu or btrend-at-maroon.tc.= umn.edu Coordinator, Characterization Facility University of Minnesota Center for Interfacial Engineering =
100 Union St SE Room 187 phone 612-624-1365 fax 612-626-7530=
In July and September of 1994, there was some disciussion of BDMA vs. DMP-30. I saved and consolidated 13 of those messages into a single document. If anyone is interested in getting a copy, send me your e-mail address and I will zip one out to you within a day or two.
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"MICROSCOPY & MICROANALYSIS 96" in Minneapolis, Minnesota, Aug.11-15, 1996
Folks, I'm aware of several programs for the Mac that simulate diffraction patterns and stereographic projections, etc. How about similar programs for the PC? Thanks in advance.
Well it may not sopund exciting to microscopists until you note that it tranlates into Noran's parent company buys Kevex's parent company! March 3rd issue of Wall Street Journal. Thermo Instrument Systems to acquire Fisons PLC's scientific instrument division. Cost $320million! Anyone know whether this covers VG too?
Just thought you guys might like to know.
-- John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu, John.F.Mansfield-at-umich.edu or jfmjfm-at-umich.edu they all reach me! URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
Hi, I know this is a little of the subject, but can anyone suggest a ftp site that has terminal emulators. We need to emulate a tektronics 4205 terminal to use our new xrd software. Any suggestions would be appreciated.
I am looking for the following pieces of used equipment either in good working order, fair condition and/or in need of minor repair:
1) glass knife breaker for LM microtomy 2) embedding oven (35-80C) 3) balance (top loading or mechanical)
Contact:
Fred A. Hayes 916-752-7712 work University of California,Davis 916-752-4701 work School of Medicine Department ofMedical Pathology; EM Lab MSIA E-mail: Davis, CA 95616 fahayes-at-ucdavis.edu
1320 Dogwood Court 916-678-6280 home Dixon, CA 95620-3227
Hi, I know this is a little off the subject, but can anyone suggest a ftp site that has terminal emulators. We need to emulate a tektronics 4205 terminal to use our new xrd software. Any suggestions would be appreciated.
Microscopy Mail {Microscopy-at-aaem.amc.anl.gov} Message-id: {01HNT9F975CY93NYWI-at-gnv.ifas.ufl.edu} MIME-version: 1.0 X-Mailer: Windows Eudora Version 1.4.4 Content-type: text/plain; charset="us-ascii" Content-transfer-encoding: 7BIT
At 08:26 AM 3/3/95 +0100, NICK SCHRYVERS wrote: } REGARDING obj. lens 100C replacement } } The objective lens and the upper and lower column parts of our 20 year old } side entry JEOL 100 C microscope need replacement. New parts, however, are } rather expensive in view of the age of the instrument so we're looking for } anyone who has some second hand spare parts for sale. Sincerely, } Nick Schryvers tel: 32-3-2180247 fax: 32-3-2180257 } EMAT, University of Antwerp } Groenenborgerlaan 171 } B-2020 ANTWERP } Belgium ********************************************************** There is a JEOL 100 C that may be headed for the scrap heap. They might be willing to give you parts. Contact Dr. Henry Aldrich, P.O. Box 110700, University of Florida, Gainesville, FL 32611 USA FAX 904-392-5922 (no e-mail) or his department chairman, Edward Hoffmann, at the same address and FAX number who gets E-mail at EMH-at-GNV.IFAS.UFL.EDU
Good Luck -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
please change my address to yi_huang-at-qmgate.anl.gov. I'd like to continue the subscription at the new address. Thank you. Yi Huang Argonne National Lab building 212/c221 building 212/c221
All instruments divisions of fisons are going to Thermo Instrument Systems including VG.
Just what we all needed. I sure service and parts are going to be even easier to get.
Mike
--------------------------------------------------------------------------- | Michael Dunlap | lab (916) 752-0284 | | Facility For Advanced Instrumentation | fax (510) 422-2282 | | University of California | mrdunlap-at-ucdavis.edu | | Davis CA, 95616 | | ===========================================================================
We have successfully used LR White resin for embedment of monolayers grown in Leighton tubes. The problems encountered were the same ones you encounter when using LR White in open molds or silicone rubber molds, or just about anything other than gelatin capsules. It's most convenient to embed coverslips in flat, open molds such as the silicone rubber type. We had such problems with this (ie excluding oxygen, penetration of the molds by the resin, etc) that we began embedding the coverslips standing upright in gelatin capsules. The long narrow coverslips of the Leighton tubes are just accommodated by a 00 gelatin capsule. This neccessitates considerably more hand trimming to get to the monolayer profile, but embedding problems are eliminated. I might also add, that once the block face is prepared for cutting, the exposed coverslip may be peeled away from the embedded cells, greatly facilitating cutting. If you do this, I would also recommend mounting the sections on a support filmed grid, as the cells will be on the edge of the section. Good Luck!
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
X-Mailer: WinNET Mail, v2.30 Message-ID: {463-at-oimag.win-uk.net} Reply-To: Software department {software-at-oimag.win-uk.net} To: microscopy-at-aaem.amc.anl.gov
The following advertisement has recently been placed in the press and may be of interest to those who read this Newsgroup who would like to work in the UK:
SOFTWARE DEVELOPMENT FOR MICROANALYSIS The Software team at Oxford Instruments Microanalysis Group develop high quality instrumentation products to satisfy customers in an international marketplace. Currently we have a number of vacancies for individuals who would enjoy the challenge of solving complex problems to generate leading edge products. If you are competent in Visual Basic, C++ or C, have a basic understanding of electronics and computer interfacing and feel you could make a strong contribution in any or all of the following areas please contact us:
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----------------------------------------------------------------- Please reply to this e-mail with the name of the person you wish to recieve it in the subject (i.e. FAO John Smith), as this is a shared e-mail address.
The following is a copy of the job posting that recently appeared on the University of Michigan Gopher Server UM GopherBlue.
The full URL is: gopher://gopher.itd.umich.edu:8888/00/acadaff/hris/gopher/Job%20Postings/Pro fessional%5CAdministrative/-at-39-at-RES%20ASSOC%20II%20ENGR%20%28Materials%20Scie nce%20%26%20Engineering%29
Job Title: RES ASSOC II ENGR Grade: 09 Min/Max $ 25,500/ 64,500 Posting Number: T-95-0826-LM Materials Science & Engineering Open Date: 03/06/1995 Close Date: 03/10/1995 Jobclass: 12871 Hours: 40.00
DUTIES: Operate and train users in the use of the lab instruments: a JEOL 200FX Analytical Electron Microscope, a JOEL 4000EX High Resolution electron Microscope, a Philips EM420 Transmission Electron Microscope, an ElectroScan E3 Environmental Scanning Electron Microscope, a Digital Instruments Scanning Force Microscope, a PHI 5400 X-ray Photoelectron Spectrometer and a PHI 600 Scanning Auger Microprobe; the successful candidate will not necessarily need to be proficient in the use of all instruments specified, but familiarity with at least four is essential; other duties include: aiding users with reduction and analysis of data recorded on lab instruments; aiding users with sample preparation equipment and preparing samples; help other lab staff maintain lab equipment; maintain lab darkroom and supplies; take part in collaborative projects with other members of the University; develop personal research projects as time permits.
DESIRED QUALIFICATIONS: Knowledge of Analytical Electron Microscopy, Scanning Transmission Electron Microscopy and High Resolution Electron Microscopy; TEM expertise should include detailed understanding of diffraction contract, defect imaging, selected area diffraction, high resolution imaging, high resolution image simulation and analysis; experience with computer programming in FORTRAN, C or Pascal.
MINIMUM QUALIFICATIONS: Master's degree or equivalent in a scientific related field; demonstrated prior related work experience; familiarity with computer systems such as Apple Macintoshes, PCs and/or UNIX workstations; working knowledge of general transmission electron microscopy and scanning electron microscopy; experience with standard data analysis for analytical techniques such as X-ray energy dispersive spectroscopy (XEDS), electron energy loss spectroscopy (EELS) and micro diffraction; good interpersonal skills.
Present Applications For This Position To The Following Office:
Ann Arbor Campus Employment Services Office Room G250 Wolverine Tower 3003 South State Street Ann Arbor, Michigan 48l09
(313) 764-6580
8 AM to 5 PM, Monday - Friday
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu, John.F.Mansfield-at-umich.edu or jfmjfm-at-umich.edu they all reach me! URL: http://www.engin.umich.edu/~jfmjfm/jfmjfm.html
Hello Barbara! I am afraid that, under the conditions you describe for fixation, significant shrinkage should be unavoidable whatever fixative would be used. I already mentioned that picric acid 'per se' in the fixing fluid does not ensure adequate preservation of acrosomial structure which is required for easy classification of spermatids (and thus of stages of the seminiferous epithelium.) What should be avoided in any case is the presence of acids usually part of most recipes for fixatives, such as acetic acid and the like. If you use '10 percent formalin', you may experience some problems because you in fact never know the actual composition of such solution: it polymerizes on the shelf and becomes also quite acidic! You should prepare a solution of phosphate buffered (pH 7.2) 4 percent formaldehyde from paraformaldehyde powder on the same day (or the previous one) that you are sacrificing the animals (you should find recipes for making such solution in any practical handbook for microscopical technique ;-)).
Fixation through immersion of the testes in the fixative may be almost acceptable for mice testes because they are small; this won't be the case for other species, even for the rat. You may try to make small (with caution!) incisions in the albuginea with a very sharp razor blade when the testes have stayed for at least an hour in the fixative, in order to improve penetration of the formaldehyde. Another trick to improve diffusion and fixation is to add from 0.5 to 1.0 percent (in volume) of DMSO (dimethylsulfoxide) to the fixing fluid (best is to combine both...)
BUT,LAST and PROBABLY _MOST_ IMPORTANT: you should realize that formaldehyde fixation, i.e. crosslinking, proceeds exceedingly slowly! Therefore, tissues should stay in the fixative at least two weeks (one year or more won't harm!) before being further processed (two weeks probably is a conservative estimate). Assessment of pathological tissues fixed for only a few hours in NBF may frequently be required for practical reasons, but don't expect such methods to result in nice preservation of testis tissue, especially if you need estimating quality of spermatogenesis!
You also may try to enhance fixation - and shorten the duration required for fixation - by performing it in the microwave oven: several cycles of no more than one minute at a time (perhaps 10 times, tissue temperatures exceeding 50 degrees celsius should be avoided), if you are indeed in a hurry and can't wait several weeks for fixation to proceed at its usual pace. If you try this, don't forget to place a water load (a becher container with about 750-1000 ml of water) besides your fixative containing flask in the oven.
That's all what I can think of. HTH Good luck John
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
I was unable to reach Nick Schryvers at his E-mail address therefore my reply is below. Perhaps others will be interested:
Nick writes: } } } The objective lens and the upper and lower column parts of our 20 year old } } } side entry JEOL 100 C microscope need replacement. New parts, however, are } } } rather expensive in view of the age of the instrument so we're looking for } } } anyone who has some second hand spare parts for sale. Sincerely, } } } Nick Schryvers tel: 32-3-2180247 fax: 32-3-2180257 } } } EMAT, University of Antwerp } } } Groenenborgerlaan 171 } } } B-2020 ANTWERP } } } Belgium } } **********************************************************
} } My reply:
There is a JEOL 100 C that may be headed for the scrap heap. They } } might be willing to give you parts. Contact Dr. Henry Aldrich, P.O. Box } } 110700, University of Florida, Gainesville, FL 32611 USA FAX 904-392-5922 } } (no e-mail) or his department chairman, Edward Hoffmann, at the same } } address and FAX number who gets E-mail at EMH-at-GNV.IFAS.UFL.EDU } } } } } } Good Luck } } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- } } Greg Erdos Phone: 904-392-1295 } } Scientific Director, ICBR EMCL } } 218 Carr Hall Fax 904-846-0251 } } University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu } } Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
I concur with Dr. Desclin's suggestions- however, I would add that pH and osmolarity of your solutions is easy to do, and can be helpful. Perfusion fixation of the testis is also a possibility. Those are my humble additions-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Tue, 7 Mar 1995, Desclin Jean wrote:
} } } Hello Barbara! } I am afraid that, under the conditions you describe for fixation, } significant shrinkage should be unavoidable whatever fixative } would be used. } I already mentioned that picric acid 'per se' in the fixing fluid } does not ensure adequate preservation of acrosomial structure which } is required for easy classification of spermatids (and thus of } stages of the seminiferous epithelium.) } What should be avoided in any case is the presence of acids usually } part of most recipes for fixatives, such as acetic acid and the like. } If you use '10 percent formalin', you may experience some problems } because you in fact never know the actual composition of such } solution: it polymerizes on the shelf and becomes also quite acidic! } You should prepare a solution of phosphate buffered (pH 7.2) 4 percent } formaldehyde from paraformaldehyde powder on the same day (or the } previous one) that you are sacrificing the animals (you should find } recipes for making such solution in any practical handbook for } microscopical technique ;-)). } } Fixation through immersion of the testes in the fixative may be } almost acceptable for mice testes because they are small; this won't } be the case for other species, even for the rat. You may try to make } small (with caution!) incisions in the albuginea with a very sharp } razor blade when the testes have stayed for at least an hour in the } fixative, in order to improve penetration of the formaldehyde. } Another trick to improve diffusion and fixation is to add from 0.5 to } 1.0 percent (in volume) of DMSO (dimethylsulfoxide) to the fixing } fluid (best is to combine both...) } } BUT,LAST and PROBABLY _MOST_ IMPORTANT: you should realize that } formaldehyde fixation, i.e. crosslinking, proceeds exceedingly } slowly! Therefore, tissues should stay in the fixative at least } two weeks (one year or more won't harm!) before being further } processed (two weeks probably is a conservative estimate). Assessment } of pathological tissues fixed for only a few hours in NBF may } frequently be required for practical reasons, but don't expect such } methods to result in nice preservation of testis tissue, especially } if you need estimating quality of spermatogenesis! } } You also may try to enhance fixation - and shorten the duration } required for fixation - by performing it in the microwave oven: several } cycles of no more than one minute at a time (perhaps 10 times, tissue } temperatures exceeding 50 degrees celsius should be avoided), if } you are indeed in a hurry and can't wait several weeks for } fixation to proceed at its usual pace. If you try this, don't forget } to place a water load (a becher container with about 750-1000 ml } of water) besides your fixative containing flask in the oven. } } That's all what I can think of. HTH } Good luck } John } } } *********************************************************** } * Jean C. Desclin (John), Associate Prof. of Histology * } * Laboratory of Histology - Faculty of Medicine * } * Brussels Free University (U.L.B.) * } * e-mail: desclinj-at-ulb.ac.be (internet) * } * snail mail: route de Lennik 808 * } * B - 1070 Brussels Belgium * } *********************************************************** }
} One of the divisions of our lab teases apart sural nerve tissue that have } been kept in a 1:1 mixture of epon:propylene oxide. The samples are very } easy to tease when the mixture is very fresh. The problem is that samples } became back-logged and were not able to be teased right away. A quick } decision had to be made, and the samples were frozen at -70C until they } could be teased. } } The nerve fibers need to be infiltrated to be teased but not polymerized } or else they become too fragile. A problem is becoming evident that after a } long period of storage, the nerve samples are being fully infiltrated } and polymerized despite being frozen (which we thought would almost stop } the polymerization process because it is largely dependant on heat). } Realizing that hind-sight is 20/20, would it have been better to leave the } accelerator out of the epoxy mixture? What affect would this have on the } quality of the resin? I understand accelerator to act as a catalyst being } a part of the polymerization reaction but not really consumed. Any } opinions to offer? } } Thanks, } } Dennis } Why do you store the fibers in resin? Isn't teasing them very messy? I teased sural nerve fibers when working for a neuropathologist. I fixed them and stored in buffer. Also I sometimes used collagenase, which made the teasing easier. } } } }
Message-Id: {MAILQUEUE-101.950307092030.384-at-vanlab.paprican.ca} To: "Griffin, Robin" {rgriffin-at-eng.uab.edu} , microscopy-at-aaem.amc.anl.gov
Hello, I have information on an "Imagist" image analysis system from Princeton Gamma Tech (PGT) which seems to be finding quite a bit of popularity. I have come across it several times in speaking to users during my hunt for an EDX system. Imagist runs on a sparc station. PGT in New Jersey, USA , can be reached at (609) 924-7310.
I have not used this system I have only seen it in use at the university here. Goodluck, Laurie
On 1 Mar, 1995 Robin Griffin wrote: } } I'm interested in finding out what the most commonly used image analysis } systems for light microscopy, materials applications. I am aware of } home-built, Beuler, Leco, and Kontron/Ibas. What else is out there and what } is used most in both University and industrial settings? } } Thanks for your help. } }
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
We are considering purchasing a scanner for TEM negatives.
1. Is this a good idea?
2. Is there a consensus on which one to buy?
Any and all input appreciated. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
A regular full time position is open in The Jackson Laboratory Biological Imaging Department - Histology Laboratory. The Jackson Laboratory is a non-profit independent laboratory founded in 1929 on the premise that the causes of cancer and other diseases could be discovered through Mammalian genetic research. The Laboratory specializes in mammalian genetics using inbred laboratory mice as model systems to study health problems such as cancer diabetes, anemia, heart disease and aging. Located on a large island in the gulf of Maine and surrounded by Acadia National Park, The Jackson Laboratory is currently undergoing a major expansion of its scientific staff and its research facilities.
Applicants with a Bachelors degree and two years related laboratory experience working with Murine specimens preferred. The position includes routine histological techniques ie paraffin embedding, single and serial sectioning and cryotomy in conjunction with Immunohistochemistry techniques, staining using heavy metals as well as other special stains.
The successful candidate must be a self starter, pay attention to detail and be able to work independently with little supervision. This individual will be responsible for providing services in support of numerous diverse research projects, must interact well with multiple users and work productively in a team environment. The position includes opportunities for advancement.
Salary range is mid to high $20,000 plus benefits and is negotiable depending on level of experience.
Interested applicants should send CV to:
Joanne Bradt Employment Specialist The Jackson Laboratory 600 Main Street Bar Harbor Maine 04609 (207) 288-3371 ext. 1281 (207) 288-3371 ext. 1082 FAX jcb-at-aretha.jax.org
Dear Greg, There are at least two types of scanner to consider depending on your application. For scanning images of the usual type with many gray levels, a CCD array will give excellent results and is very fast; however, for quantita- ting ED patterns, nothing beats a scanner with a small illuminating spot. The problem with a CCD array for ED is that the highest intensities are in areas with very low transmission surrounded by areas with high transmission, so in- ternal reflections in the CCD lens, etc. will give erronious values for the intensities. Any of the high-resolution CCD arrays will work for images, since the changes in contrast are not so abrupt. Perkin-Elmer and Optronics are two names for small-illuminating-spot scanners, and doubtless there are others. We had an Eikonix (linear CCD array) at one time, but were not happy with it. Good luck. Yours, Bill Tivol
Here's a little information that I just received from polaroid that should be of interest to the microscopy community. I'd be interested in hearing any comments anyone (particularly any Polaroid reps.) might care to share with the BBS.
Recently we have been noticing that the type 55 film we are purchasing was arriving with a relatively short period before expiration. We were questioning if our supplier might have trying to unload older film on us. So I just called up Polaroid an asked them what their lead time from manufacture to expiration date was, and I was informed that for type 55 and type 665 (B&W P/N film) the expiration lead time is only 9 months.
If you allow say 2-5 months from manufacture to shipping to distributors to shipping to users (which seems very reasonable for such commercial retail) this leaves only 4-7 months before expiration! This short of an expiration date does not lend itself at all to buying in larger quantities in order to obtain any sort of a price break.
I do not know how much extension can be obtained by storage at say 4 C (May be someone out there does know). But we seem to start having problems with film only a couple of months after the expiration date even after cold storage for a few months.
You might want to consider this short time to expiration before you go and stock up on polaroid film.
Richard E. Edelmann Electron Microscopy Facility Supervisor Miami University, Oxford, OH 45056 Ph: 513-529-5712 E-mail: edelmare-at-muohio.edu
Although the journal Microscopy Research and Technique has a large editorial board, we cannot cover every research area that involves microscopy. Therefore, I would like to invite those of you who are interested in serving as ad hoc referees for articles submitted to the journal to send, by e-mail, your name, mailing (regular mail) address, phone number, fax number, and research interests using key words such as biology, materials science, pathology, microanalysis, stereology, magnetic domain, ceramics, immunocytochemistry, kidney, etc.
John E. Johnson, Jr., Ph.D. Editor-in-Chief, Microscopy Research and Technique
We are considering purchasing a scanner for TEM negatives.
1. Is this a good idea?
2. Is there a consensus on which one to buy?
Any and all input appreciated. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
Ed, We have had 3 different tissue processors in the past 6 years, 3 LKB's, one Lynx, and 3 RMC's model 4189. LKB no longer manufactures a system so I will forego 'ripping' their machine. We purchased a Lynx system about four years ago. We found the same problem that W.L. Steffens had with the QC of the expendibles. We lost tissue due to improper sealing of the specimen chambers. Also the side 'openings' are relatively large and we therefore do not run tissue smaller than 0.5 mm3 in these holders. Extreme care must be used when removing tissue from the holders because tissue has a tendency to adhere to the lid of the holder making it possible to mix tissues when using the multiwell chambers. The unit itself though has been extremely reliable. About 10 months ago we purchased 3 RMC processors. These have proven not to be as reliable as the Lynx (we have had temperature and some programming problems). We therefore have had the opportunity to test RMC's service department which to date has been able to get us up and running with minimal down time. The big advantage of the RMC units are the specimen chambers which seal better and hold smaller pieces of tissue. Currently we use only the RMC units for tissue processing (approximately 3000 samples annually). The Lynx system has been modified and is used 3-4 days (and evenings) a week to immunolabel grids for our IEM program. Hope this helps.
{jon charlesworth} Electron Microscopy Facility Mayo Clinic
We are considering purchasing a scanner for TEM negatives.
1. Is this a good idea?
2. Is there a consensus on which one to buy?
Any and all input appreciated. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 904-392-1295 Scientific Director, ICBR EMCL 218 Carr Hall Fax 904-846-0251 University of Florida E-mail: gwerdos-at-gnv,ifas.ufl.edu Gainesville, FL 32611
} noticing that the type 55 film ... with a relatively short period before } expiration } } But we seem to start } having problems with film only a couple of months after the } expiration date even after cold storage for a few months.
Please forgive the above condenstation :-}
For the last 15 years we have stored Polaroid type 55 film in a freezer without harm. This appears to negate the expiration date. We usually allow the film to thaw out completely before opening the sealed package, but sometime this process is rushed a bit and then, more often than not we have problems.
We have found that to be the situation for years, but we order it by the case and refrigerate it (not freezing). It seems to be good for at least a year past expiration and we have had some film that was several years old still work. I suspect that there could be some degradation, but for our routine SEM and light microscopy, it hasn't been a problem. Maybe Polaroid has done some testing?
***************************************************************** Michael L. Boucher Sr. Boucher-at-TCRCA.USBM.GOV Geology-Mineralogy/Chemistry Labs Ph 612-725-4614 Twin Cities Research Center Fax 612-725-4527 U.S. Bureau of Mines Center 725-4500 Department of Interior 5629 Minnehaha Avenue South Minneapolis, MN 55417-3099 U.S.A. *****************************************************************
I am going to be attempting to thin section sediment samples containing quartz and want the thin sections to be approx. 50-70nm in thickness for TEM analysis. I have to buy a diamond knife and would much appreciate input on the blade angle to choose. A representative at Polysciences told me I should go to a 55 degree knife instead of a 45, but I was wondering if I would be sacrificing much of the ability to cut sections down to 50nm. The quartz grains are approx. 200 um in diameter. Is there a particular manufacturer I should get the knife from? What is an appropriate cutting speed? Can I expect the knife to last only a VERY short time? I would appreciate any suggestions and advice. My address is: chswartz-at-MIT.edu
Does anyone on the listserver know where (prefereably in the U.S.) that I can get Carbon Putty. I'm down to the last of what I had that someone picked up at a conference in Germany a few years ago. It is basically a mixture of some kind of soft wax and carbon and is a very good specimen mounting material since there is no drying or out-gassing. Ted Pella doesn't seem to have it so maybe someone out there knows about this product and can save me a hunt.
} I do not know how much extension can be obtained by storage at say } 4 C (May be someone out there does know). But we seem to start } having problems with film only a couple of months after the } expiration date even after cold storage for a few months. } } Richard E. Edelmann
We are just finishing our last few boxes of September 1994 Type 55 film (stored in a refrigerator). It still seems to be OK.
Arthur Day, Electron Microscopy Group Ansto Advanced Materials Program Phone: 61-2-717-3457 PMB 1, Menai (Sydney), NSW, 2234 Fax: 61-2-543-7179 Australia Email: ard-at-atom.ansto.gov.au
------------------- tn8502.asc follows -------------------- Tracor 8502 users: Help with image convert to tiff
We operate a Tracor Northern (Noran) 8502 Image Analyzer and have been trying to export images to a VAX via KERMIT with no success. Has anyone done this sort of image file conversion and transfer successfully?
Background: The Tracor software can convert an image file from TN's format (.IMG) to a .TIF file (type 4, version 42). We can successfully transfer this TIFF image file, via KERMIT, to or VAX (ie. the file appears and has the appropriate size). When we try to open this TIFF image (using a PC on our LAN), we always fail regardless of the software we have employed (e.g. with Photoshop, HiJack Pro).
If you have any ideas, please contact. I have lots of additional contextual information to add to this. Thank you!
Eric Kokko Electron Microscopy and Image Analysis Agriculture and Agri-Food Canada Lethbridge Research Centre P.O. Box 3000, Lethbridge, Alberta CANADA T1J 4B1
Phone 403-327-4591 (Voice 367) FAX 403-382-3156 INTERNET kokko-at-em.agr.ca
Someone has just given me all you could ever want to know about this product. It is available from: Bal-Tec Products, Inc. 984 Southford Road P.O. Box 1221 Middlebury, CT 06762; (800) 875-3713, FAX (203) 598-3658
It is called "Leit-C-Plast; Cat. # B 801014075 Current price is $70.00 for 15g
Other properties: -high electric conductivity -long-life plasticity -high vacuum resistance -sufficient adhesive power -low sample contamination -no disturbing ED-x-ray lines -Resistance R~100 kOhm mm2/m Thanks for saving me the hunt---Jerry
"Mung II" is not carbon putty, but is very much like it: electrically conducting, a good thermal conductor, and very low vapor pressure. We use Mung in our ion mill, but in the SEMs most people prefer clips or carbon tape. You can buy mung from
Comonwealth Scientific Corp. 500 Pendleton St. Alexandria, VA 22314 703-548-0800
$160 for 5 gr.
_______________________________________________________________ Michael Rooks 607-255-2329 voice National Nanofabrication Facility 607-255-8601 fax at Cornell University rooks-at-nnf.cornell.edu Knight Laboratory Ithaca, NY 14853 USA
_______________________________________________________________ Michael Rooks 607-255-2329 voice National Nanofabrication Facility 607-255-8601 fax at Cornell University rooks-at-nnf.cornell.edu Knight Laboratory Ithaca, NY 14853 USA
You are asking about a product called "Leit-C-Plast", which is found on page 46 of the current SPI Supplies "SourceBook" as SPI #5057-AB. It is a highly conductive adhesive plastic and is designed for mounting large specimengo a very long way. I hope this information will be useful to you.
Charles A. Garber, Ph. D. PRESIDENT SPI SUPPLIES PO BOX 656 West Chester, PA 19380
"Richard E. Edelmann" {EDELMARE-at-CASMAIL.MUOHIO.EDU}
Richard Edelmann wrote " I'd be } interested in hearing any comments anyone (particularly any Polaroid } reps.) might care to share with the BBS. } } Recently we have been noticing that the type 55 film we are } purchasing was arriving with a relatively short period before } expiration. We were questioning if our supplier might have trying to } unload older film on us. So I just called up Polaroid an asked them } what their lead time from manufacture to expiration date was, and I } was informed that for type 55 and type 665 (B&W P/N film) the } expiration lead time is only 9 months. " } I have used Polaroid 665,667,55 for nearly 20 years with no problems of any continuing significance (except maybe cost !!). We have always purchased in bulk (large boxes of 50 boxes) and stored the material in a standard refrigerator, not freezing it. Usual practice would be to allow the boxes to equilibrate at room temperature before use but sometimes usage rate prevented this. The sensitivity does appear to change with temperature, but we have nver bothereded to investigate in detail. In the usual situation, we have not expereinced any problems and have both negs and prints which are old and still very good quality. Cleanliness of the holder and specially the rollers is critical to performance and they are cleaned after EVERY cartridge is exhausted and before the next one is in place. cheers jjm Professor John J. Millar Head, Department of Applied Physics Electron Microscope Unit RMIT, GPO Box 2476V, MELBOURNE AUSTRALIA 3001 +613 660 2602 fax 613 660 3837 email jjmill-at-rmit.edu.au
RE: Ultrathin sectioning material containing quartz
We have had some amount of experience in our own laboratory with the thin sectioning of these kinds of samples for TEM characterization.
There are the obvious trade offs, that is, th that a) the price is much cheaper and b) you are using the "normal" narrow angle (e.g. 45 deg.) knives and therefore you don't have to worry about compression effects.
While the ability to cut sections is somewhat related to the experience of the one doing the sectioning (and it is also an art), so long as you are patient, you should be able to do it.
If you wanted to send us one of your samples, we could produce sections for you, and essentially determine the exact conditions under which you too could easily get sections. There would be a fee of course for this, however, you would know for sure, one way or another, whether sections could or could not be made of your material,and if they could, then you would also know the exact cutting conditions.
Another important consideration would be the choice of embedding resins and we would recommend for quartz particles our SPI # 2660 SPI-PON 812 resin kit since this particular resin seems to be the most user friendly in terms of polymerizing to a hardness for which sections can be made of the embedding quartz particles.
You have also asked a question that touches on the expected longevity of the diamond knife when cutting these kinds of materials. You sort of have to think about it as an analog to the mileage expected from a pair of tires. You can buy imported tires or you can buy domestic ones, however, what really counts is how you drive your car. Be gentle, and the knife will last longer. Be abusive, and it will have a shorter life time. Obviously, cutting quartz particles is going to be "hard" on the knife. You can extend the longevity be making the particles smaller before embedding. You can get good sections faster, and therefore also less knife wear, by curing the SPI-Pon 812 resin harder right away, putting less wear and tear on the knife to get to that point.
I hope this information has been helpful to you. Let me know if you have any other questions.
Charles A. Garber SPI Supplies PO Box 656 West Chester, PA 19380 USA
The Swiss Society of Optics and Electron Microscopy, the French Society of Electron Microscopy and the Belgian Society of Microscopy have decided to hold a joint meeting in 1995. This conference will be held at the "Ecole Polytechnique Federale de Lausanne" (EPFL), Lausanne, Switzerland, from June 26 to 30, 1995. It will include two tutorial days and a three-days conference which will be mainly focused on the recent progresses in TEM and scanning probe microscopies for materials science and biological fields.
********** SCIENTIFIC PROGRAMME **********
Monday 26 and Tuesday 27 TUTORIALS, with practical training, for groups of 6 to 20 people.
- Cryo-microscopy of vitrified specimens: theory and practice (Jacques Dubochet) - Practical aspects of the methodology of image analysis in oncology (Ricardo Laurini) - TEM image simulation of crystalline materials (Pierre Stadelmann) - Electron holography (Conradin Beeli) - Analytical microscopy with a field emission TEM (Klaus Leifer) - TEM sample preparation in materials science (Daniele Laub) - Applications of large angle convergent beam electron diffraction techniques (LACBED) (Jean-Paul Morniroli)
Evening Tuesday 27 Welcome ceremony and conference opening lecture.
"Perception du monde exterieur par les systemes vivants" by Yves de Ribaupierre, University of Lausanne.
Wednesday 28, Thursday 29 and Friday 30 - Conference
- New microscopies: STM, AFM, SNOM, confocal, ultra-sound, ... (A. Engel, D. Pohl)
- Energy filtered images and electron spectroscopy (C. Colliex, D. Bazett-Jones)
- Life in extreme environment (F. Gaill)
**** BIOLOGY-PATHOLOGY SYMPOSIA ****
- Fertilization and early embryogenesis in mammals (J. E. Flechon)
- Freeze substitution, microscopy of frozen hydrated samples (J. Dubochet, S. Fakan)
- Image analysis and cancer diagnosis (R. Laurini)
- High-resolution electron microscopy of biological specimens (E. Delain, M. Muller)
- Cytoskeleton (G. Gabbiani)
- New applications of scanning probe microscopies (M. Robert-Nicoud)
**** PHYSICS-MATERIALS SYMPOSIA ****
- Electron holography (C. Beeli, D. van Dyck)
- New progress on CBED/LACBED (J. P. Morniroli, J. Steeds)
- High-Resolution microscopy of aperiodic structures (J. van Landuyt, P.Stadelmann)
- New applications of analytical microscopy: filtered images, EDS, EELS (C. Colliex, C. Humphreys)
- New applications of scanning probe microscopies (D. Pohl, D. Courjon)
**** Commercial exibition ****
A substantial commercial exhibition of scientific equipment will also be held during the conference. For information, please contact G. Peter, EPFL-CIME CH 1015 Lausanne Fax: +41 [21] 693 44 01, email: peter-at-cime.epfl.ch
********** REGISTRATION **********
The symposia will consist essentially of a small number of lectures (mainly invited, with some selected from the conference abstracts) and of poster sessions. The presentations may be given in English, or in any languages of the three national societies. However, certain parts of the presentation should be in English, at least abstracts, figure captions, transparencies... Discussion sessions will be devoted to a few hot points emerging from the poster presentations.
The registration fees for the conference, including attendance at the symposia and the abstract booklet, are 250 CHF (non-members), 200 CHF (members) and 100 CHF (students and lab technicians). The registration fees for the tutorial days will be communicated in the final circular. A maximum of 100 CHF/course is expected. A few grants have been founded by the national societies in order to encourage the participation of young scientists and technicians
********** GENERAL INFORMATION **********
The conference will be held at the Ecole Polytechnique Federale de Lausanne (EPFL), located approximately 4 km from the city centre, near (500 m) the idyllic scenery of lake Leman. Hotel rooms in different categories (from "Formule 1" to ****) have been reserved by the tourist office for the organization committee, with prices ranging from 25 (F1, 3 persons per room) to 230 CHF. A final hotel booking form will be included with the registration form.
Excursions for participants and accompanying persons are planned: e.g. a tour to the beautiful region of "la Gruyere" famous for its cheese, to a wine cellar on the coast of lake Leman !! ... These excursions will be organized in small groups of 20 to 30 persons according to their preferences.
********** ORGANIZING COMMITTEES **********
Honor committee: Walter Bollmann, Alain Gautier, Eduard Kellenberger, Bernard Vittoz
International Scientific Committee: M. Deschuyteneer (SmithKline Beecham, B), J. Dubochet (Uni. Lausanne, CH), A. Engel (Uni. Basel, CH), J. Gunter (Uni. Zurich, CH), D. Hernandez-Verdun (Inst. Jacques Monod, F), J. Lecomte-Beckers (Uni. Liege, B), J.-P. Morniroli (Uni. Lille, F), R. Portier (ENSCP, F), M. Praet (Uni. Gent, B), D. Schryvers (Uni. Antwerpen, B), P. Stadelmann (EPFL Lausanne, CH), D. Thomas (Uni. Rennes, F)
Local Scientific Committee: M. Campiche (Uni. Lausanne), J. Dubochet (Uni. Lausanne), S. Fakan (Uni. Lausanne), R. Gotthardt (EPFL Lausanne), R. Laurini (Uni. Lausanne), P. Stadelmann (EPFL Lausanne)
Local Organizing Committee: P.A. Buffat (Chairman), R. Rouquier (Secretary), C. Beeli, F. Bobard, B. Garoni, P.-H. Jouneau, D. Laub, G. Peter, B. Senior, P. Stadelmann
I am looking for information about software to read .bmp or TIFF files using a Mac. We have a Targa+ video capture board to capture images from the SEM onto a 486-66 PC. I would like to be able to import them over e-mail to a Mac Quadra 650 and paste into a Word document. Word will import the image, but there seem to be a decrease in resolution and the file size is reduced from 650K to 375K when it is converted to the Mac. I would like to here from other users who are capturing images as we are just getting the system set up. Thanks,
John Giles Honeywell Space Systems jegiles-at-space.honeywell.com (813)539-2270 phone (813)539-3630 Fax
} I do not know how much extension can be obtained by storage at say } 4 C (May be someone out there does know). But we seem to start } having problems with film only a couple of months after the } expiration date even after cold storage for a few months. } } Richard E. Edelmann
I had a problem of inside sticky #52 Polaroid film, which will expire on Sept. 1995, so that run out of our film. I found a box of old #52 film which expired on Aug. 1992 left in a drew (at room temperature) and tried to use it. To my surprise, it worked fine. For my experiences, the problem of Polaroid does not really related to the expiration date. Instead, I found a lot of problems are caused by storage. I have used a lot of good expired films and a lot of fresh films as well for 5 years.
Ya Chen Integrated Microscopy Resource Madison, WI 53706 USA Email: ychen-at-macc.wisc.edu
} We are in the process of purchasing a new printer and are looking at the } Tektronix Phaser II SDX, Kodak ColorEase PS, and Kodak 8600. Samples } provided by } Kodak and Tektronix look great. I have heard that the Mitsubishi S3600-32U } is a } great printer but cannot get any useful information, prices, or samples from } Mitsubishi or local dealers. I am curious about the Fujix 3000. Could you let } me know how or where to contact Fujix, since I have never heard of them.
You can contact Ron Saltzman, at Fujix Electronic Imaging Group at 800-736-3600 ext. 8282 (voice mail). If he's no longer with that group, you might try the general numbers, 914-789-8100 or 800755-3854, to get to a sales rep. They also advertise in some of the photo lab trade magazines.
Good luck. They are really worth a look.
John chandler-at-lamar.ColoState.EDU Fort Collins CO
Thanks to those who have helped me locate sources where I could find a material matching the description I gave for what I called "carbon putty."
Since posting the first source given to me (Bal-Tec) yesterday, I have received a few more. So, for those other subscribers who asked me to post any info. I got, here are the other sources.
Marivac LTD./Hailfax, N.S./Canada/(800)565-5821/Cat.# AS508-3/ $66.50 Canadian 15g (Approx. $50.00 American)
SGL-Carbon/Niagara Falls, N.Y./P.O.Box 667/(716)236-2859/ This company makes carbon epoxy material that sets-up for permanent attachments. It is not a soft putty after it sets but it is highly conductive and may be of some use for other purposes.
Hello All, I have a user who was told to try Zamboni Fix on their tissue. Could some kind soul tell us what it is and how to make it. I know what and Zamboni is in relation to ice rinks and I can even drive one (it's a real hoot!!) but I've never heard of the fix... TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
Ed Calomeni wrote: } I am in the info gathering stage of purschasing a tissue processor for EM } samples. } I have info on the Reichert-Lynx unit and the one made by RMC. How } do you like } these units? Comments on advantages and disadvantages are } welcome.
} Thanks in advance
} Ed Calomeni } Medical College of Ohio } Toledo, OH
We have a Lynx tissue processor which we are now very happy with. There were a few teething problems when it was first installed (the main control board and the motor control unit had to be replaced) but these problems I believe have now been sorted out at the factory and shouldn't apply to new instruments now - it was some three years ago when we bought our one. The only other problem we had was with voltage spikes from the mains. This problem was eliminated by putting an isolating transformer between the mains and the processor. We have not experienced any problems with the samples mixing upon opening up the vials (someone -sorry I forget who- mentioned this in an earlier reply). Infiltration of resin seems to be fine, something I was a little sceptical of initially, given the small holes in some of the basket types. There is a good choice of different specimen baskets anyway and I have always found one suitable for whatever specimens I happen to be processing. Hope this is helpful for making your decision. Regards, RE
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
Message-Id: {199503092054.PAA03408-at-alchemy.chem.utoronto.ca} X-Sender: pmarkiew-at-alchemy.chem.utoronto.ca (Unverified) X-Mailer: Windows Eudora Version 1.4.4 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Fellow Microscopists,
This is a general announcement which should be of interest to atomic force microscopists.
We are making the latest version of our deconvolution program MIDAS95 available for free on the Internet. The program is designed to work under Windows on Nanoscope files. It is being released as a beta version, meaning the program lacks a certain polish. The program works properly for both the Nano II and III, although testing on the latter system has been limited.
Through deconvolution, one can account for the volume occupied by the tip in the AFM images obtained. By eliminating the tip effect, the result is often a truer representation of the sample topography. Deconvolution also allows for the in situ measurement of the tip geometry. This 3D data file of the tip can be used as a check of the tip's integrity or for improvement of other AFM images. Further details are given in the README.TXT file.
The program can be accessed by the following: ftp surfturf.chem.utoronto.ca login: anonymous password: yourname-at-location Please use your E-mail address for the password. We wish to keep a record of the users of this program and notify them of any problems or updates.
The public directory has three files of interest here. One is the README.TXT file if you want to know what is in MIDAS.ZIP without having to download it. MIDAS.ZIP contains several files, including some AFM files to see how deconvolution is done and some TIFF files for viewing. Fan-xing Wei and Dr. Dan Thomas of the University of Guelph have modified our program so that it works for Burleigh Instruments. This program is available in BURL.ZIP.
If you have any difficulty, please be patient as our server allows only one user at a time and tends to lock up when doing calculations.
The best way to get bitmap and/or TIFF images into your Mac is with the NIH Image software (via ftp: anonymous-at-zippy.nimh.nih.gov directory /pub/nih- image). It can handle either format, but you will probably have to go through the IMPORT diaolog, not the OPEN dialog. Look at the documentation (also on Zippy) to guide you through. As long as there is a full valid bitmap in the image file, NIH Image can import the image and allow you to save it as a useful file. We have done a lot of image file format trouble-shooting for "standard" formats written by multiple vendors on multiple platforms around here using that capability.
A plug here: NIH Image has a nearly infinite performance to price ratio - it is free once you are on the internet. It also outperforms a whole lot of commercial stuff on any computer platform up to about the $2000 price point and the support system is unbelievably good, considering the annual maintenance non- fee. 8-).
Two hurdles are: getting the file onto your Mac (which it sounds like you are already doing) then getting a good image into Word. I do the latter by pasting the image from NIH Image into Word, then shift-shrinking the image to about 50%. The original paste goes in at 72 dpi, the shift-shrink gives the final image resolution of 144 dpi which, with a decent gray-scale printer, gives you very acceptable print quality. By the way, I __STRONGLY__ advise against using the image editor in Word. I have found it unreliable in addition to the fact that it reduces your grayscale image to 16 levels. This could explain the drop in file size (the editor converts the 8-bit image to 4-bit). A similar result would happen if you started with a 16-bit TIFF and wound up with 8-bit.
Hope this helps.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Message-Id: {9503092328.AA21558-at-riker.ml.wpafb.af.mil} To: WALCKSD:ML:WPAFB cc: Lloydpf:ML:WPAFB Subj: ultrathinsectioning material containing quartz In-Reply-To: Message from WALCKSD:ML:WPAFB of 3-8-95 ----------------------------------------------------------------------- Scott,
I would recommend that the knife of choice be a Diatome knife. The angle would depend on how hard the sediment is. If the sediment is very hard, then yes I too suggest a 55x knife. Using a knife with this angle still allows one to obtain sections on the order of 70nm. If the sediment is not that hard, then a 45x knife will work also. I generally use a 55x knife for all materials that I cut except for polymers, where I use a 45x knife.
I hope this helps!
Pam
---------------------- Replied Message Body ---------------------- To: LLOYDPF Subj: ultrathinsectioning material containing quartz Forwarded: Message from {chswartz-at-MIT.EDU}:ddn:wpafb of 3-8-95 ------------------------------------------------------------------
--------------------- Forwarded Message Body --------------------- To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: ultrathinsectioning material containing quartz Orig-Author: {chswartz-at-MIT.EDU}:ddn:wpafb ----------------------------------------------------------- I am going to be attempting to thin section sediment samples containing quartz and want the thin sections to be approx. 50-70nm in thickness for TEM analysis. I have to buy a diamond knife and would much appreciate input on the blade angle to choose. A representative at Polysciences told me I should go to a 55 degree knife instead of a 45, but I was wondering if I would be sacrificing much of the ability to cut sections down to 50nm. The quartz grains are approx. 200 um in diameter. Is there a particular manufacturer I should get the knife from? What is an appropriate cutting speed? Can I expect the knife to last only a VERY short time? I would appreciate any suggestions and advice. My address is: chswartz-at-MIT.edu
I'd appreciate a copy of the BDMA vs DMP-30 responses.
Regards
Mike Gregory
} In July and September of 1994, there was some disciussion of BDMA vs. DMP-30. I } saved and consolidated 13 of those messages into a single document. If anyone is } interested in getting a copy, send me your e-mail address and I will zip one out } to you within a day or two. } } -- } } Gib Ahlstrand, MMS Newsletter Editor } Electron Optical Facility, University of Minnesota, Dept. Plant Pathology } 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 } 612-625-9728 FAX, giba-at-puccini.crl.umn.edu } } "MICROSCOPY & MICROANALYSIS 96" in Minneapolis, Minnesota, Aug.11-15, 1996 } }
} The best way to get bitmap and/or TIFF images into your Mac is with the NIH } Image software (via ftp: anonymous-at-zippy.nimh.nih.gov directory /pub/nih- } image). It can handle either format, but you will probably have to go through } the IMPORT diaolog, not the OPEN dialog. Look at the documentation (also on } Zippy) to guide you through. As long as there is a full valid bitmap in the } image file, NIH Image can import the image and allow you to save it as a useful } file.
As a follow-up, we use NIH Image for our image processing and also import BMP and TIFF files from PC capture systems. NIH Image will import TIFF directly using 'OPEN', and from our experience BMPs should be IMPORTed using CUSTOM, setting OFFSET to 1760 and chsing the relevant bitmap size.
I hope this is of use
Doug
+------------------------------------+ | Dr Douglas Arrell | | Mechanical Performance and Joining | | Institute for Advanced Materials | | 1755 ZG Petten | | Netherlands | | {ARRELL-at-JRC.NL} | | Tel. (+31) 2246 5287 | | Fax (+31) 2246 1917 | +------------------------------------+
A short message for users of (or people interested in) 'S-SIMPLY' (SHRLI-SIMulation and diSPLaY of TEM & HRTEM images, for PCs) : the freeware version, available on FTP.UNIV-LYON1.FR (/pub/dos/HRTEM) has been significantly updated, and new features are available (graphical displays, a "Make Interface" tool,...). With best regards,
______________________________________________
Thierry EPICIER GEMPPM-502, INSA de Lyon, 69621 VILLEURBANNE, France tel : (33) 72 43 84 94 (83 85) FAX : (33) 72 43 85 28 Email : EPICIER-at-CISMSUN.UNIV-LYON1.FR
Two days ago I posted a Cat.# and price for Leit-C-Plast from Bal-Tec that was given to me incorrectly. The correct Cat.# is B801014077 and the cost is $40.00 for 15g. This is now the best price I've found.
I recently purchased a Pictography 3000 printer after a extensive comparison of printer systems available on the market. The printer is attached to the network (Token ring, novell) via a pc, and is transparent configured: all pc's in the network can reach the printer. The printer is used for production of hard-copies of image files generated by various techniques: CSLM, VEM, cryo-SEM, FESEM (+EDS Noran SUN), TEM (Gatan SSC, Mac), SPM, IA (SEMPER).
We are quit satisfied.
E-mail: Marcel.Paques-at-2488PAS.URLNL.Sprint.Com Unilever research Laboratory Vlaardingen The Netherlands
Zamboni fix is an EM version of the classical LM Bouin's fix. Zamboni's original abstract didn't contain any details about how to make it up. The details are in: Stefanini et al., 1967, Nature 216:173.
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School {akc-at-umich.edu}
------------------------------------------
On Thu, 9 Mar 1995, Michael Stanley wrote:
} Hello All, } I have a user who was told to try Zamboni Fix on their tissue. } Could some kind soul tell us what it is and how to make it. I know what } and Zamboni is in relation to ice rinks and I can even drive one (it's a } real hoot!!) but I've never heard of the fix... } TIA, } } C. Michael Stanley, Ph.D. } Coordinator, Associate Director } Molecular Cytology Core Facility } Molecular Biology Program } 2 Tucker Hall } University of Missouri-Columbia } Columbia, MO. 65211 } (314) 882-4895 } fax= 314-882-0123 } } }
I have recently been shown a research report in BioTechniques, Vol 13, No 3. (1992) by Reed, J.A., Manahan, L.J., Park, C-S, and Brigati, D.JL describing an immunocytochemical method based upon capillary action. This was using the "MicroProbe" marketed by Fisher Scientific, Pittsburgh, PA.
My question is, has anyone used this system? If so what are its advantages and/or disadvantages . Also could you please give me a contact address, FAX number or E-Mail for Fisher Scientific.
Message-Id: {9503101815.AA2900-at-pho018.sb.com} To: images1 {images1-at-biosci.mbp.missouri.edu} , microscopy {microscopy-at-aaem.amc.anl.gov}
In response to this question:
Hello All, I have a user who was told to try Zamboni Fix on their tissue. Could some kind soul tell us what it is and how to make it. I know what and Zamboni is in relation to ice rinks and I can even drive one (it's a real hoot!!) but I've never heard of the fix... TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
I think that the fixative you're referring to is cited in this paper: Zamboni, L. and DeMartino, C., 1967. Buffered picric acid-formaldehyde: A new, rapid fixative for electron microscopy.
We came across Zamboni's fix when we were perfusion-fixing rat testes a couple of years ago. However, it didn't preserve Leydig cell structure as well as some of the other fixatives we tried.
Hope this info helps. (I want to hear more about driving a Zamboni!)
Bev Maleeff SmithKline Beecham Pharmaceuticals Toxicology-US, UE 0462 709 Swedeland Road King of Prussia, PA 19406 610/270-7987 610/270-7202 fax e-mail: maleeffbe-at-sb.com
The 30th annual meeting of the SouthEastern Microscopy Society (SEMS) will be held on May 17-19, 1995, at the Omni Hotel in Atlanta, GA. Scheduled invited speakers and their titles are: 1) Nestor Zaluzec, Argonne National Laboratory, Integrating Computers, Microscopy, and Microanalysis; 2) Phil Russell, North Carolina State University, Instrumentation and Application of Scanned Probe Microscopy; 3) Larry Peterson, GBI, Forensic Microscopy; 4) Terence Mitchell, Los Alamos National Laboratories, Experience with Field Emission on an SEM, STEM, and TEM; and 5) Jay Jerome, Bowman Gray School of Medicine of Wake Forest University, Exploring Ultrastructure with Quantitative 3-D Intermediate Voltage Electron Microscopy. Also on the program are pre-meeting workshops and tours, contributed papers and posters, the RUSKA student competition, and commercial exhibits. Social events are the Wednesday night Exhibitor's Mixer and the Awards Banquet, which will be held Thursday night at the Fernbank Museum of Natural History
For further information, contact Janet H. Woodward, SEMS '95 Program Chair, at (912)-945- 3152, FAX (912)-945-3155.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
My goodness I got the message several times. Also the one I sent came back to me several times. I guess the Government wants to make sure evereything gets through.
Too bad about the cuts. Frank Skelton told me about Van Closing also I guess the Farm got hit.
PCI is selling well particularly in the USA and some to Japan.
I have full internet access now. PPP with every application which is Freeware and Shareware. All windows based and relatively easy to use.
Our site in the office is working under a private account, but we haven't got the domain name yet. It will be NSCTOR. Anyway I will let you know when it comes through.
David Cockayne asked: } } WITH REGARD TO THE QUESTION ABOUT TEACHING OPTICS, CAN ANYONE ADVISE ME OF WHAT SOFTWARE PACKAGES ARE AVAILABLE FOR TEACHING ELECTRON OPTICS?
At the same time a similar question was posted on the Microscopy list (by someone different).
I would be delighted if anyone can update the following list of teaching software (in the area of microscopy) known to me:
The Institute of Materials (UK) distribute:
The Transmission Electron Microscope by Goodhew The Scanning Electron Microscope by Humphreys (John) Electron Diffraction by Goodhew The Stereographic Projection by Humphreys Analysis in the Electron Microscope by Goodhew, Humphreys and Cliff X-ray Photoelectron Spectroscopy by Watts X-ray Auger and Laser emission by Goodhew
These are all IBM PC DOS versions, available from Institute of Materials, 1 Carlton House Terrace, London, SW1Y 5DB, fax +44(0)171 839 2078 The MATTER project is currently working on greatly-improved Windows and Mac versions of most of these topics: Completion of the first two topics (TEM and SEM) is scheduled for the end of 1995. All the above are aimed firmly at teaching, not at research applications.
There is also an SEM simulation available which originated in Australia but which is supposed to be downloadable from Nestor Zaluzec's Microscopy Bulletin Board. However we have consistently failed to transfer it. Has anyone in the UK succeeded?
I also saw recently a very full simulation of a light microscope. This is aimed at biologists and was written (and is distributed) by Maurice Smith at: mol-at-molcol.demon.co.uk
If you can add to this list please reply to both lists [ microscopy and materials-l ]
Thanks
Peter
PS 120+ members of the materials list now!
------------------------------------------------------------------ ----- Professor Peter J Goodhew, Department of Materials Science & Engineering University of Liverpool LIVERPOOL Fax (44) (0)151 794 4675 L69 3BX, UK Tel (44) (0)151 794 4665 (secretary Debra) ------------------------------------------------------------------ ----- inter alia: Director of the MATTER project for educational software ------------------------------------------------------------------ -----
STUDENT COMPETITION TO BE HELD AT THE UNIVERSITY OF WISCONSIN WHITEWATER, WISCONSIN Ambrose Health Center, Room 162 Friday, March 24, 1995 PROGRAM: 1:00 PM: Dr. Ursula K. Charaf, Senior Research Scientist, Johnson Wax: BLOW IT UP! Applied Industrial Microscopy 1:45 PM: Student Presentations 3:00 PM: Dr. Robert Cardell, University of Cincinnatti Medical Center, MSA/LAS Sponsored Presidential Speaker: Modern Microscopical Approaches to Biomedical Problems 4:00 PM: Reception and Award Presentations
If you are planning to come, telephone or E-mail registrations are required by March 22. If you are planning to present your research, please send your abstract to: Dr. Lance Urven University of Wisconsin- Whitewater 800 West Main Whitewater, WI 53190 urvenl-at-uwwvax.uww.edu PHONE 414-472-5132 Reception provided by Oxford Instruments Inc., Microanalysis Group.:
Don- 1) the thin film coatings on the polycarbonate are very tightly held proprietary secrets. try some EDS or WDS. 2) try SEM to determine where these "pits" reside. 3) the "trick" is to plunge the CD into liquid nitrogen (30 sec), then give it a shot with a hammer. it shears at the interface
we are investigating the CD's by AFM and SEM here in our lab, keep in touch about further results?
-Mike
On Sun, 12 Mar 1995, Chernoff wrote:
} I am seeking advice regarding specimen preparation. I wish to } examine the physical pits created when a CD-Recordable disk is } written. I would like to understand: } - the physical arrangement of thin film coatings on the } polycarbonate substrate, i.e. what materials are stacked up, and } what their typical thicknesses are. } - what layer contains the physical pits. } - how to expose that layer for examination by SEM or AFM. } } Thanks for your help. } } DON CHERNOFF 317-251-1364 } ADVANCED SURFACE MICROSCOPY FAX: 317-254-8690 } 6009 KNYGHTON RD. E-MAIL: echernof-at-ucs.indiana.edu } INDIANAPOLIS IN 46220 Toll free: 800-374-8557 } } }
Where could I findout what was the fair market value for a JEOL 1200ex II 5 years old. Basic TEM no Scanning EDX or stuff like that. Who has the blue book??
I'VE BEEN ASKED TO ADDRESS A GROUP OF STUDENTS (LATE HIGH SCHOOL AND EARLY COLLEGE) A LOCAL COLLEGE SCIENCE CAREER DAY. I AM PLANNING ON USING A VARIETY OF SEM MICROGRAPHS TO ILLUSTRATE THE APPLICATIONS OF THE SCANNING ELECTRON MICROSCOPE TO MANY SCIENTIFIC AREAS. IF ANYONE HAS ANY PC-FORMAT IMAGES THAT THE WOULD BE WILLLING TO E-MAIL TO ME, I WOULD BE VERY GRATEFUL.
There really isn't anything equivalent to a "blue" book, since there are just not enough instruments of this type getting sold in order to get any kind of meaningful sales statistics. However, there are basically three different types of prices discussed:
a) Selling price of a used instrument from the OEM
b) Selling price from a third party service company
c) Offering price from a former owner
If you want to find out (a), the best person would be Robert Santorelli at JEOL in Peabody, MA Ph; (800) 343-6766.
For (b), you should contact Mr. Clark Houghton, Secondary Images, Winchester, OH FAX: (513) 927-5557.
For (c), you should contact again, either Mr. Houghton (above) or Mr. James Nicolino, X-ray Optics, Florida, Ph: (904) 646-3069 FAX: (904) 565-1897. Mr. Nicolino services wave length dispersive microprobe systems but has a good understanding of what different instruments fetch when sold by an owner, free of any guarantee or warranty, and such sales are generally on an "as is, where is" basis.
Hope this information will be useful to you.
Charles A. Garber SPI Supplies West Chester, PA 19380 USA
To: goodhew-at-liverpool.ac.uk Order #4457898 From: GVKM07A Date: 03/13/95 11:46 PM
Prof. Peter Goodhew
Hello! I think that the soft ware you are thinking of is called the emTutorial system, and it is indeed produced in Australia. There are actually two different system programs, emTUTOR and semTUTOR, and they have been designed to be used as a self-instruction tool, an instruction aid for class labs and for lecture demonstrations.
Also I think the commercial producers of these really outstanding software products would be quite unhappy if it should turn out that their proprietary software was distributed without any benefit to them. I could be wrong about that, but if I was a betting person, that would be my opinion.
The product can be ordered from SPI Supplies as SPI #09000-AB and has a regular price of $345. By coincidence, we have been running a "special" and until April 15, the price is $250.
There is another new product from the same Australian firm called PARFOCAL, which is a graphics file conversion program for confocal microscopy and image analysis. The regular price is $325 but this product is not current on any kind of special sale.
If you provide a FAX number, we can FAX you additional information.
Charles A. Garber SPI SUPPLIES (USA) PO Box 656 West Chester, PA 19380 USA
Ph: (800) 242-4774 Toll free in USA only (610) 436-5400 Regular phone number (800) 526-6562 Toll free in Canada only - rings in USA
FAX: (610) 436-5755 1-800-55-7780 Toll free FAX from Ireland only 0800-44-0873 Toll free FAX from New Zealand 0800-89-3066 Toll free FAX from UK
I have received the following information from the organizers and wanted to publicize it to anyone interested in attending or participating:
First International Conference on Electron Microscopy and Advances in REsearch in Different Fields of Science
September 2-4, 1995 Ismailia-ETAPT
Sponsored by Electron Microscopy Center Suez Canal University Ismailia EGYPT
Special Topics of the Conference: 1] Role of EM in diagnostic virology 2] Role of EM in diagnosis of tumors cytology and urinary stones 3] Role of EM in ultrastructure pathology of the lung (non neoplastic conditions) 4] X-ray microanalysis: Applications particularly metallurgical, mineralogical, and biological 5] Scanning EM of plants, animal, insects, and mineral material 6] Study of biological macromolecules from their characteristic electron diffraction patterns 7] Skin pathology by EM 8] Morphological identification of antigens by EM 9] Different low temperature methods for biological EM 10] Safety measures and maintenance needed for EM
There will be an equipment exhibition in conjunction with this meeting.
Registration for foreigners will be US $150 inclusive of full board during the time of the meeting.
For further information, contact the organizer:
Prof. Dr. Khalifa Ibrahim Khalifa Electron Microscope Center Suez Canal University Ismailia EGYPT
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To: GVKM07A From: Peter Goodhew Date: 03/13/95 06:01 AM
David Cockayne asked: } } WITH REGARD TO THE QUESTION ABOUT TEACHING OPTICS, CAN ANYONE ADVISE ME OF WHAT SOFTWARE PACKAGES ARE AVAILABLE FOR TEACHING ELECTRON OPTICS?
At the same time a similar question was posted on the Microscopy list (by someone different).
I would be delighted if anyone can update the following list of teaching software (in the area of microscopy) known to me:
The Institute of Materials (UK) distribute:
The Transmission Electron Microscope by Goodhew The Scanning Electron Microscope by Humphreys (John) Electron Diffraction by Goodhew The Stereographic Projection by Humphreys Analysis in the Electron Microscope by Goodhew, Humphreys and Cliff X-ray Photoelectron Spectroscopy by Watts X-ray Auger and Laser emission by Goodhew
These are all IBM PC DOS versions, available from Institute of Materials, 1 Carlton House Terrace, London, SW1Y 5DB, fax +44(0)171 839 2078 The MATTER project is currently working on greatly-improved Windows and Mac versions of most of these topics: Completion of the first two topics (TEM and SEM) is scheduled for the end of 1995. All the above are aimed firmly at teaching, not at research applications.
There is also an SEM simulation available which originated in Australia but which is supposed to be downloadable from Nestor Zaluzec's Microscopy Bulletin Board. However we have consistently failed to transfer it. Has anyone in the UK succeeded?
I also saw recently a very full simulation of a light microscope. This is aimed at biologists and was written (and is distributed) by Maurice Smith at: mol-at-molcol.demon.co.uk
If you can add to this list please reply to both lists [ microscopy and materials-l ]
Thanks
Peter
PS 120+ members of the materials list now!
------------------------------------------------------------ ------ ----- Professor Peter J Goodhew, Department of Materials Science & Engineering University of Liverpool LIVERPOOL Fax (44) (0)151 794 4675 L69 3BX, UK Tel (44) (0)151 794 4665 (secretary Debra) ------------------------------------------------------------ ------ ----- inter alia: Director of the MATTER project for educational software ------------------------------------------------------------ ------ -----
-------- Original message header follows -------- From goodhew-at-liverpool.ac.uk Mon Mar 13 06:01:17 1995 [PIM 3.2-342.56] Received: from AAEM.AMC.ANL.GOV (aaem.amc.anl.gov [146.139.72.3]) by inetgate.prodigy.com (8.6.10/8.6.9) with SMTP id GAA64180 for {GVKM07A-at-mail.prodigy.com} ; Mon, 13 Mar 1995 06:01:17 -0500
To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: AFM Learning Curve ------------------------------------------------------------------
One of our researchers is looking into buying an AFM, primarily for studying latex coatings. As a microscopist (PLM,SEM,EDX) in the analytical group, I'm interested in the broader applications we might find for our entire product line.
These are my questions:
1.) What is the "typical" learning curve for the AFM for non- microscopists vs. microscopists?
2.) How useful is it for one to have access to other types of microscopies to correlate with AFM results?
3.) Does the usual rule of "fewer operators, less downtime" apply to any greater or lesser degree with the AFM?
I look forward to any thoughts and experiences you can share.
Dave Stadden DRStadde+aCENTRAL%ARMSTRONG-at-MCIMAIL.COM Phone 717-396-5109 FAX 717-396-5865
I am the author of "Virtual SEM", an earlier version of which called "SUPERSEM" is available through FTP from the EMMPDL library.
"Virtual SEM 1.2" is an update of this earlier package and is much enhanced. I'm a bit short of time but the recent discussion (Prof. Goodhew, and Chuck Garber - regards to both) I think is in part mistaking my software for that available commercially through SPI and others.
The latest version will be passed to Nestor and John Mansfield in about a fortnight at the Scanning meeting and I hope they will see fit to update the old version. It is public domain and comments are most appreciated. To summarise it is a MAC product utilising real images controlled through a "SEM console" simulation. It includes tutorial information and two self assessing tests, the results from which are automatically stored on file. We have been using it for 18 months and feedback has been good.
The drawback is that it is now around 35 meg in size and I expect it to reach 40 meg soon. It only needs 4 meg of ram to run on, for example, a MAC IIsi or better. I developed most of it on a powerbook 180C.
The size is a problem for FTP and as noted in earlier versions I can supply it on a multisession CD. To do this I am charging US$75 to cover writing and postal costs only. I will upgrade to later versions and hope to be able to at postal cost, if I have to hire a bod to do it then would have to cover the time but cost would still be minimal. We also have a virtual EDS in development and plan to fully integrate the two.
I will try and answer any queries. Those that have contacted me earlier, I have not lost your requests, am working my way thru as fast as I can - please be patient.
Brendon Brendon J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-9-380-2739 fax 61-9-380-1087
I am an Associate Professor at The Rockefeller University interested in obtaining access to the microscopy listserver. A colleague told me that this would be a good medium to advertise postdoctoral and technical positions that may be available in my laboratory.
Please let me know if I need to submit any more information. My address is: William A. Muller, MD, PhD Box 176 The Rockefeller University 1230 York Avenue New York, NY 10021 Phone (212) 327-8104 Fax (212) 327-8875
I have heard rumors that there will be an international microbeam analysis meeting in Australia early in 1996. Does anyone know if this is so; and if it is, do you have any information about when and where it will be?
I'm staining spinal cord and medulla's neurons from rats' fetuses, 10 days old, with the following primaries antibodies: Rabbit anti Glycine 1:2500, 1:4000; Rabbit anti GABA 1:100, 1:200; Rabbit anti GAD 1:300, 1:500; and anti-synaptophysin 1:150, 1:300. I'm using FITC 1:100 or Rhodamine 1:100 as my secondary antibodies. I keep getting crossover between FITC and Rhodamine despite the different dilutions of the primary antibodies I have used. Does anyone has any suggestion? Thanks in advance,
Ciprian Almonte Research assistant Medical College of Pennsylvania Dept. of Anatomy and Neurobiology 3200 Henry Ave. Philadelphia, PA 19129 Voice: (215) 842-4081
We've been trying to measure "integrated density" of some fluorescent images and found significant differences in the numbers calculated in v. 1.53 and 1.57. We have 1.53 on a quadra 950 and 1.57 on an 8100. Working with the same image, 1.53 returns numbers approximately 2,000,000 and 1.57 gives us numbers about 400. The mean intensity, Std dev, background, number of pixels...are identical in both versions. LUTs are also the same in both versions. These are TIFF images acquired on a DOS machine. I'm not sure if it's related to the integrated density difference, but in 1.57 we can "open" the TIFF files, while in 1.53 we have to "import" the TIFF file.
Is there something obvious I'm overlooking? Any suggestions are appreciated. Thanks
I'm staining spinal cord and medulla's neurons from rats' fetuses, 10 days old, with the following primaries antibodies: Rabbit anti Glycine 1:2500, 1:4000; Rabbit anti GABA 1:100, 1:200; Rabbit anti GAD 1:300, 1:500; and anti-synaptophysin 1:150, 1:300. I'm using FITC 1:100 or Rhodamine 1:100 as my secondary antibodies. I keep getting crossover between FITC and Rhodamine despite the different dilutions of the primary antibodies I have used. Does anyone has any suggestion? Thanks in advance,
Ciprian Almonte Research assistant Medical College of Pennsylvania Dept. of Anatomy and Neurobiology 3200 Henry Ave. Philadelphia, PA 19129 Voice: (215) 842-4081
A general posting since I've had a bunch of questions all about the same thing.
Yes the EMMPDL/MASLIB/PD Shareware libraries are avaiable via anonymous FTP. The site is
WWW.AMC.ANL.GOV (= 146.139.72.10 )
Login via standard FTP protocols with Username=Anonymous Password=Your Email Address
but please note that you CANNOT access these files using a WWW Browsing program like Mosiac, Netscape or any other, even though they have FTP capabilities. You must use a standard FTP protocol client program. which accesses the conventional FTP ports.
The reason for this is that the although WWW server and the FTP server are both on the same Computer (WWW.AMC.ANL.GOV=146.139.72.10) they reside on two different disk partitions which are NOT linked. The FTP server looks in one place and the WWW server another. When you login to the WWW site, your browser (client) program has only access to the disk space assigned to the WWW server hence it (the WWW program) will NEVER see the FTP files and vice versa! The reasons for this are a combination of security and convenience on my part, but with the explosion of the WWW I've created yet another headache for myself.
So for all of you that have tried and failed this is the most common reason. If you don't understand this call me next week sometime and I'll explain, or better yet come to the Telecommunications Tutorial which I'll be giving at the August MSA meeting.
In retrospect I should have put them on different machines with different DN's and then the problem would not have happened, but the damage is done and until I get a chance to come up for air it will have to stay that way. There is yet another alternative which is to define an alias on the DNS for the FTP server. If I can figure out the right person to contact I'll try that route but be guarenteed it will not be soon.
Just a reminder..... Abstracts/Papers for the August MSA meeting are due March 15th i.e. probably the day you receive/read this message.
If anyone has information on the Microscopy Meeting in August could you please send information on contacts for the meeting.
Also, I am looking to attend regional meetings and would love to find a calendar of meetings and places with contacts. I would be attending for training and exibiting.
Thank You all!!!
P.S. all that are interested about the LaserMaster ImagePrinter 1800 DPI the "NEW" price is $ 6,995.00 and an upgrade from a 60 mhz processor to a 100 mhz. WOW, 1K off plus upgrade.
Greg Begin - LaserMaster Imaging Specialist in Scientific/Medical Imaging
Just read the posting by Ciprian Almonte concerning crossover between FITC and Rhodamine and had a couple of questions about what he is doing. What are your FITC and Rhodamine linked too--goat anti-rabbit??? Are you trying to do double labelling??? Or is it that you are getting fluorescence of the FITC at the rhodamine settings?? More information is required to help. Yours Mark Elliott, PhD UBC-Pulmonary Research Laboratory, St. Paul's Hospital Vancouver BC Canada
You need to "fine tune" your microscope filters. Do you perhaps have an older Olympus?? Call your local microscope rep and ask for the correct filters to narrow the band pass of the dichroic filters. Grace
I think this may have been discussed by this group earlier but at the time we were not worried about it so I didn't keep a record of the responses. One of our tech's asked me to post this: We are looking for a plastic medium which can replace Epon in Biological TEM. In our lab we regularly use Epon 812 for its good stainability of membranes. Recently however, we have just too many samples to infiltrate and Epon is just too viscous for multiple tissue transfers. The result is extensive numbers of labour hours. Please let us know if you have succcessfully worked with any plastic that has equal stainability of the membranes but is less viscous. Thanks Mark Elliott UBC-Pulmonary Research Laboratory St. Paul's Hospital Vancouver BC Canada
} Just read the posting by Ciprian Almonte concerning crossover between } FITC and Rhodamine and had a couple of questions about what he is } doing. What are your FITC and Rhodamine linked too--goat } anti-rabbit??? Are you trying to do double labelling??? Or is it } that you are getting fluorescence of the FITC at the rhodamine } settings?? More information is required to help. } Yours } Mark Elliott, PhD } UBC-Pulmonary Research Laboratory, } St. Paul's Hospital } Vancouver BC Canada
FITC and Rhodamine are linked to anti-rabbit, and I'm planning to do double labelling. My major problem is that I'm getting flourescence of the FITC at the rhodamine settings and viceversa. Thanks, to all of those that have replied so far. I appreciate all their suggestions. Ciprian
Ciprian Almonte Research assistant Medical College of Pennsylvania Dept. of Anatomy and Neurobiology 3200 Henry Ave. Philadelphia, PA 19129 Voice: (215) 842-4081 E-mail almonte-at-medcolpa.edu
Try using more selective excitation filters (narrower band pass). Another suggestion is to use a "Red Cut" Filter. This greatly supresses the secondary peak that is associated with FITC. Contact you local microscope supplier or Omega Optical at (802) 254-2690. Good Luck,
Hello all, We have a user wanting to look at the inside of Polycaprolactone Microcapsules in the SEM. We are not wanting to do sections for the TEM. I have tried to fracture the microcapsules in liquid N2, after embedding them in resin, but they dont fracture through the capsule, only around it. Also, I have tried to section the block face of the capsules in resin, but resin seems to infiltrate into the capsule and so no internal structures can be seen. These capsules are between 100 microns - 20 microns in diameter, and have a melting point around 65oC. Has anyone tried this before and had success? The references I have contain no details about preparation of their specimens. Any help would be appreciated.
G'day Subscribers (actually in it's now about 11 pm in Chicago)
Here's the info that was posted about the IUMAS meeting that I dug out of the Archives. You will have to check with Dave Williams of Lehigh (dbw1-at-lehigh.edu) if you want further details.
Also the WWW site is also starting to get fairly busy, lately we've been averaging ~ 250 connections/day. So if it appears slow, consider yourself forwarned.
The FTP server crashed sometime on Monday Evening. So if anyone tried to get to the libraries, it was a useless battle. As of a few minutes ago it was back up and running.
Why does this always happen just before deadlines for MSA abstracts?
It must be the Ides of March syndrome, but then again maybe it was a Leprechaun.
MICROBEAM ANALYSIS SOCIETY STUDENT SCHOLARSHIPS TO MAS-1995 (Breckenridge, CO) and IUMAS-1 (Sydney Australia)
All students (and faculty) involved in microanalysis-related research, should note a remarkable opportunity for travel to two forthcoming microanalysis conferences. The Microbeam Analysis society is offering student scholarships to the 29th Annual MAS meeting in Breckenridge CO (August 6-11, 1995). Student papers must be submitted, in standard MAS-MSA format to Dr. E. Etz, NIST, Division 837, Bldg. 222, room Q-113, Gaithersburg, MD 20899 by March 15, 1995. The best submitted papers will be awarded funds towards attending the annual meeting in Breckenridge. Any more information about the program can be obtained from Dr. Etz at etz-at-gapnet.nist.gov
What makes this scholarship offer extraordinary is that the best three papers given by student scholarship winners at the Breckenridge meeting will be awarded a minimum of $500 towards the cost of attending the 1st International Union of Microbeam Analysis Societies Conference in Sydney, Australia, February 5-9, 1996. These scholarships are only open to student members of MAS, and student application forms for MAS are available in past issues of Microbeam Analysis, the MAS journal. Student membership is a great bargain at $2.50, and doesn't require that the advisor be a MAS member - although if you aren't, I ask that you consider joining.
I will mail you more information with appropriate details about the meetings and the paper format etc., but if you have any immediate questions, please don't hesitate to contact me by email (dbw1-at-lehigh.edu).
Can anyone out there point me in the right direction for finding information on the health risks associated with epoxy resin dust produced by jewler's saws, jig saws, dremel tools, files, sand paper, etc.. Yes, I have looked at the various MSDS for the resins but they haven't provided much real world information (I at least haven't run into the problem of a tanker truck spillage of NSA).
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Biological Science Building Miami University, Oxford, OH 45056 Ph: 513-529-5712 E-mail: edelmare-at-muohio.edu
We use double stick tape to fracture the Microcapsules. On a SEM stub we mount an stip of tape. On tape that is covering the stub we add a small amount of dried microcapsules. Using an exta piece of tape cover the tape and capsules and rip off. This will fracture many of the microcapsule so that you will be able to look inside and outside many of them. Coat the stubs and view in the SEM.
Good luck
Mike
--------------------------------------------------------------------------- | Michael Dunlap | lab (916) 752-0284 | | Facility For Advanced Instrumentation | fax (510) 422-2282 | | University of California | mrdunlap-at-ucdavis.edu | | Davis CA, 95616 | | ===========================================================================
I have done many TEM preparations on cell monolayers, using tissue culture dishes and/or glass coverslips. There is a lab here that would like to do light microscopy and TEM on a single cell in a monolayer. I assume this can be done, if I had a coverslip that was etched with a marker grid. This marked grid pattern would have to show on the epoxy face, after the coverslip was separated from the embedding media. Can anyone tell me if there are glass or plastic coverslips that have some type of etched grid pattern that could be used in this way and where they could be ordered in the U.S.
I would prefer working with a plastic/tissue culture type of coverslip, as they are easier to remove before sectioning.
thanks Louisa Howard Ripple Electron Micrscope Facility Dartmouth College Hanover, N.H 03755 Louisa.Howard-at-Dartmouth.edu
I have done many TEM preparations on cell monolayers, using tissue culture dishes and/or glass coverslips. There is a lab here that would like to do light microscopy and TEM on a single cell in a monolayer. I assume this can be done, if I had a coverslip that was etched with a marker grid. This marked grid pattern would have to show on the epoxy face, after the coverslip was separated from the embedding media. Can anyone tell me if there are glass or plastic coverslips that have some type of etched grid pattern that could be used in this way and where they could be ordered in the U.S.
I would prefer working with a plastic/tissue culture type of coverslip, as they are easier to remove before sectioning.
thanks Louisa Howard Ripple Electron Micrscope Facility Dartmouth College Hanover, N.H 03755 Louisa.Howard-at-Dartmouth.edu
I have done many TEM preparations on cell monolayers, using tissue culture dishes and/or glass coverslips. There is a lab here that would like to do light microscopy and TEM on a single cell in a monolayer. I assume this can be done, if I had a coverslip that was etched with a marker grid. This marked grid pattern would have to show on the epoxy face, after the coverslip was separated from the embedding media. Can anyone tell me if there are glass or plastic coverslips that have some type of etched grid pattern that could be used in this way and where they could be ordered in the U.S.
I would prefer working with a plastic/tissue culture type of coverslip, as they are easier to remove before sectioning.
Thanks.
Louisa Howard Ripple Electron Micrscope Facility Dartmouth College Hanover, N.H 03755 Louisa.Howard-at-Dartmouth.edu
If you are going to look at cells in a light microscope, it is a poor idea to use plastic coverslips. You will not get a good image in Differential Interference Contrast or Polarized light...as the cover slip may be strained, and so forth. N.Allen
Hello Microscopists, Although I can't help Richard Edelmann with his question regarding resin dust except to say that we do use dust masks when sawing or cutting blocks; I would like to pose a question closely related to this. How many of you have your curing ovens vented to a hood? We have a VERY low volumn of resins being cured at any given time and have sought, somewhat unsuccessfully, information that would help us determine at what level it would be necessary to do this. As Richard said, the MSDS sheets don't provide much real world information. I did call one of the EM suppliers (I don't recall which one now), and was told that venting to a hood would be necessary only in an industrial situation. I would be very interested in hearing your opinions and how you handle this problem. I would also be willing to summarize any responses and post that to the list.
Thank you very much, Sandra Zane
Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
We use glass gridded coverslips for tracking cells. You can get them from Bellco Glass, Inc. P.O. Box 340 Edrudo Road Vineland, New Jersey 08360 Cat. # 1916 -92525 Etched gridded coverslips 1 case approx. $30.00.
Patty Jansma Tel:602-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
On 16 Mar 1995, Louisa Howard wrote:
} I have done many TEM preparations on cell monolayers, using tissue culture } dishes and/or glass coverslips. There is a lab here that would like to do light } microscopy and TEM on a single cell in a monolayer. I assume this can be done, } if I had a coverslip that was etched with a marker grid. This marked grid } pattern would have to show on the epoxy face, after the coverslip was separated } from the embedding media. Can anyone tell me if there are glass or plastic } coverslips that have some type of etched grid pattern that could be used in } this way and where they could be ordered in the U.S. } } I would prefer working with a plastic/tissue culture type of coverslip, as } they are easier to remove before sectioning. } } Thanks. } } Louisa Howard } Ripple Electron Micrscope Facility } Dartmouth College } Hanover, N.H 03755 } Louisa.Howard-at-Dartmouth.edu
Hello Microscopists, Although I can't help Richard Edelmann with his question regarding resin dust except to say that we do use dust masks when sawing or cutting blocks; I would like to pose a question closely related to this. How many of you have your curing ovens vented to a hood? We have a VERY low volumn of resins being cured at any given time and have sought, somewhat unsuccessfully, information that would help us determine at what level it would be necessary to do this. As Richard said, the MSDS sheets don't provide much real world information. I did call one of the EM suppliers (I don't recall which one now), and was told that venting to a hood would be necessary only in an industrial situation. I would be very interested in hearing your opinions and how you handle this problem. I would also be willing to summarize any responses and post that to the list.
Thank you very much, Sandra Zane
Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
Glass coverslips etched with an alphanumeric grid are available from Bellco Glass, Inc. (800-257-7043). I have successfully grown cells in culture on these coverslips for LM and SEM analysis after coating the coverslips with either poly-lysine or collagen. Identified cells are relatively easy to re-identify when going from one system to the other. I have not embedded cells grown on these coverslips, so I do not know if the grid will show up on the surface of the plastic when the coverslip is removed.
Jeff Thompson, Ph.D. Director, Electron Microscope and Image Analysis Center California State University San Bernardino, CA 92407 jthompso-at-wiley.csusb.edu
I must confess I'm a bit perplexed by Z.Q. Liu's question about x-sections of Nano-tubes. If they are truely "nanometer" in size why would you attempt to x-section them? Are they actually larger? All the EM I've seen about nanotubes did not involve x-section, but simple direct imaging.
Okay so let me ask the dumb question. How big are they really?
What is the appropriate way to digitize TEM negatives for HRTEM work at resolution of approx.0.23 nm? Please give approximate cost of the digitizing hardware when possible. Leszek Kepinski
***************************************************************** Leszek Kepinski Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 937, 50-950 Wroclaw, Poland tel:(4871) 350 21 ext.153 fax:(4871) 44 10 29 email: kepinski-at-highscreen.int.pan.wroc.pl *****************************************************************
} I have been asked to analyse some micas for Rb and from a } preliminary examination it looks like I should use the L alpha } line - except that the Si K alpha is rather close making the } Rb sit on the slope to the right of it. } } Anyone have any advice on backgrounds etc? } } I am using TAP xtals, 25 kV,55 nA with a counting time of } 60 seconds for peak and background - is this reasonable? } } Machine is Cameca SX50.
25kV, 55 nA sounds pretty high for routine mica analyses, though it might be OK to analyse the Rb at this condition after analysing the major elements. At 25kV, you could use the Rb KA line on an LIF crystal and avoid the Si overlap. The counting times sound reasonable, though you may have to increase them if you are really interested in "trace" analysis of low levels of Rb.
I suspect that you will always obtain "false" Rb when analysing Rb LA on TAP in the presence of high Si in your sample. The Si KA peak is about -750 steps from the Rb LA line (and about +200 from the Rb LB line). This is far enough that most of the Si peak will be resolved, but there will probably be some tail effect of the Si under the Rb LA peak. You can check for this be analysing SiO2. This will give you the worst case Si interference for silicates. You could continue this process and draw up a working line (curve) of wt.% Si vs. wt.% measured Rb by measuring various silicates with differing Si contents and no Rb. You could then use this curve to correct for your analyses of Rb on your unknowns.
The SX50 software should also give you the chance to use a background "slope" instead of having to take background counts at both +bkgd and -bkgd positions. The "slope" on the older CAMECA MBX software was defined as (bkdg+ + bkgd-)/(2*bkgd+). Thus, a "flat" background would have a slope of 1.000, while a background that increases with lower sin(theta) will have a slope } 1.000. Using a background slope will not solve the tail effect of the Si under the Rb LA, but may help you in using the Rb KA line on LIF, where the bkgd- position may be out of range for the spectrometer.
Good luck!
Carl
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2005 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA ----------------------------------------------------- (313) 936-1550 (voice) **** Next time, **** (313) 763-4690 (FAX) *** take the *** chender-at-umich.edu (e-mail) **** train! **** -----------------------------------------------------
Dear Nestor, John Cowley gave me reprints of some of his recent papers in which there are images of nanotubes which are on the order of 10 nm across (O.D.). He and his co-workers have found different structures at various places in the nanotubes, so I can see why someone might wish to section them (although it is hard to imagine how this can be done without perturbing (destroying?) the structures). John would likely send you reprints if you asked him. Yours, Bill Tivol
Dear Leszek, In order to retain the original resolution (0.23 nm), the digitization should have a pixel size corresponding to no more than half that resolution. That is, if the magnification of the negative is 100,000x, the pixel size must be ~10 micrometers. This can be done on (among other brands) a Perkin-Elmer flatbed microdensitometer (cost ~ $100,000). A much cheaper alternative is a CCD camera. By arranging to image the negative with the CCD so that the pixels are ~10 micrometers you can get the required resolution. This means that a 1000*1000 CCD array will look at a 1 cm**2 area. This can be done at a cost of ~$50,000 (maybe less--I'm no expert here). You also save scan time with the CCD; however, quantitation of intensities--especially for ED patterns etc. where there are regions of very high OD surrounded be regions of low OD-- is much worse with CCD than with a flatbed scanner. Good luck. Yours, Bill Tivol
Subject: Time: 8:16 AM OFFICE MEMO nanotube x-sect. Date: 3/17/95
Can standard TEM images normal to the axis of the nanotube determine unambiguously the cross sectional shape of tubes (i.e. whether the tubes are facetted, circular cylinders, oval-shaped, hollow or filled with amorphous material, etc?). We think there are 2 ways to get this information: direct imaging of cross-sections or electron holography (in which case you don't need cross-sections). The problem with cross-sections is the potential development of artifacts during the preparation process. We suggest strongly to use electron holography (see for example: J. Mater. Sci., 29 (1994) 5612-5614 (this is on Pd particles, but the principle is the same). Also see our paper which includes nanotubes in the Proceedings of the 1st Int'l Workshop on Electron Holography, Elsevier, in press.
Bill Tivol said in his email that "the digitization should have a pixel size corresponding to no more than half that resolution". I think that this is a typo: you need at least 2 sampling points for a given wave. Therefore if one wants to quantify 0.23nm spacings in HREM the smallest sampling is half this (Nyquist limit) and } 6 samplings is more standard.
I've repeated the original question below, in case someone's missed the thread. I suspect that Liu is trying to make cross-sections to characterize growth mechanisms. Nanotubes, like asbestos fibers, are easily examined in plan-view, since their high aspect ratio virtually guarantees that they will lie flat on a film with the long axis parallel to the film surface. Most of the pictures that I have seen claiming to be nanutubes in cross-section had no supporting evidence that they were anything but an essentially spherically symmetric "bucky onion."
In order to get a true nanotube cross section, one will first probably have to anneal the mass to remove all lesser fullerenes, including the onions. Then break the mass up and embed in a hard epoxy. Although ultramicrotomy isn't a bad idea, there is no real way to align the nanotubes and I'm not certain what effect this would have on cutting. It is probably better just to sandwich between silicon and do a standard cross-section by mechanical polishing, dimpling, and ion-milling. It is going to be hard to prevent rapid milling of the epoxy as opposed to the tubes, so it is important to mechanically thin as far as possible before ion-milling (hence, the silicon as an optical guide).
That's my 2 cents worth.
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu ---------- Forwarded message ----------
Regarding the recent debate on negative scanning: The Ektron scanner is {$30K, and gives a highly linear image up to 4096 x 4096 pixels. It will permit accurate density corrections to be done for best quantitative results (see Voelkl, et al., Ultramicroscopy 55 (1994) 75-89, for all you could ever want to know about scanning negatives and density corrections etc.)
L.D. Marks is correct--I meant a pixel size corresponding to twice the reso- lution, or half the size. I always seem to have trouble writing about things for which less is better :-). Yours, Bill Tivol
Microscopical Society of Canada 22nd Annual Meeting
The MSC Executive and the Local Organising Committee cordially invite you to attend and participate in the 22nd Annual Meeting of the Microscopical Society of Canada. This meeting will be held in the University Centre Building, University of Ottawa, Ottawa, Ontario, Canada, June 4-7, 1995. A varied and interesting scientific program has been planned and will consist of a combination of interdisciplinary symposia presented by speakers from around the world, separate physical and biological symposia, oral and poster presentations, workshops on TEM specimen preparation of materials and cryo- SEM specimen preparation and X-ray analysis, and commercial exhibits.
Local Organising Committee
Jim Corbett (Chairman, Secretary, University of Ottawa Liaison) John McCaffrey (Treasurer, Space Management, Social Program, Accommodation) Jeff Fraser (Commercial Exhibits, Social Program, Accommodation) Graham Carpenter (Scientific Program Chair, Materials) Larry Arsenault (Scientific Program Chair, Biology) Peter Sewell (Corporate Liaison, Commercial Exhibits) Kamal Botros (Scientific Program) Louise Weaver (Scientific Program) Paula Allan-Wojtas (Registration) Shea Miller (Registration)
DEADLINE FOR RECEIPT OF ABSTRACTS: March 31, 1995
DEADLINE FOR PRE-REGISTRATION: May 1, 1995
For further information contact:
Program: Registration:
Jim Corbett Shea Miller or Paula Allan-Wojtas Department of Physics Centre for Food and Animal Research University of Waterloo Agriculture and Agri-Food Canada Waterloo, Ontario Room 2016, K.W. Neatby Bldg. Canada, N2L 3G1 Central Experimental Farm Ottawa, Ontario, Canada, K1A 0C6 Tel: (519) 885-1211 Tel: (613) 957-4347, ext. 7908 (Shea), 7970 (Paula) Fax: (519) 746-8115 Fax: (613) 943-2353 e-mail: corbett-at-physics.watstar.uwaterloo.ca e-mail: millers-at-ncccot.agr.ca allanwojtasp-at-ncccot.agr.ca
PRELIMINARY MEETING PROGRAM
INTERDISCIPLINARY SYMPOSIUM ADVANCES IN MICROSCOPY
J. Watson (Michigan State University) Events in science and microscopy J. Hillier (Princeton University) Early developments in EM A. Eades (University of Illinois) Electron microscopes of the future O. Krivanek (Gatan Inc) Nanoscale elemental and chemical mapping M. Hansen (FEI Inc.) A comparison of LaB6 and CeB6 thermionic cathodes I.A. Rauf (Queen's University) STM, AFM, CTEM and STEM imaging of polycrystalline tin-doped indium oxide thin films H.J. Kreuzer (Dalhousie University) Lensless low-voltage electron holography
MATERIALS SCIENCES
D. Dorset (University of Buffalo) Direct structure determination by electron crystallography A. Eades (University of Illinois) RHEED: progress towards the extraction of quantitative information P. Midgley (University of Bristol) Electron diffraction for structure determination D. McCombe (McMaster University) Scanning tunnelling microscopy of semiconductor surfaces and interfaces D. Perovic (University of Toronto) Direct imaging of semiconductor dopants by EM M. Loretto (University of Birmingham) Analytical TEM of advanced materials G. Weatherly (McMaster University) Microstructures of SiC-reinforced Mg casting alloys C. Hansen (Queen's University) The application of environmental SEM to studies of concrete
BIOLOGICAL SCIENCES
W. Chiu (Baylor College of Medicine) Cryo-imaging P. Ottensmeyer (University of Toronto) Imaging and computer analysis H. Hagler (University of Texas) Calcium imaging P. Ingram (Research Triangle Inst) Elemental mapping P. Echlin (Cambridge University) Cryo-methods
WORKSHOPS
TEM specimen preparation for Materials Science Topics include: Ion-beam sputtering Cleaving Ultramicrotomy Cross sections, etc.
Cryo-SEM specimen preparation and X-ray analysis Topics include: Fracturing and cryo-planing for morphometric and elemental analysis Cryo-conductive coating Standards for frozen hydrated specimens Light element detection and quantification
Satellite Workshop (June 8, Central Experimental Farm, Ottawa) Applications of Microscopy in Agricultural Research Topics include: Preparative techniques 3-D view of the cell Microtubules Soils in agricultural use Electron microscopy in dairy research Entomology Image analysis-seed quality Image analysis of SDS-PAGE gels Spore development Immunogold labelling Food microstructure
Remember, if the camera is on the ragged edge of 8 bit performance and you want to add gain for more sensitivity, even the smallest amount of gain might put you in the 7 bit range if you we not there already. If you have 10 bit capability on your board why not use a 10 bit camera. * Also a camera specification of 60dB is not a 10 bit camera only an 8. * Also there are many types of noise in a sensor besides thermal noise. Cooling a sensor will reduce (thermal noise only) and allow you to integrate longer. I would double check the impression some make that when a 50db sensor is cooled it offers a better s/n of 10 or 12 bits vs just 7. The reduction of one type of noise (thermal) is all that cooling provides. * Other types of sensor noise besides dark current include photon, nyquest, shot, gain noise, and flicker noise etc. * The signal to noise of the sensor at room temperature is a good measure of what you can expect. ** Look at the quantum efficiency at the nm range needed. ** look at the spectral response of the sensor. ** There are different grades of sensors available relative to dead pixels and gray level variation on other manufacturers cameras. *Digital RS-422 eliminates pixel jitter for sub-pixel accuracy if it is needs versus the genlock action between the digitizer board and the camera as they try to lock to each other in the RS-170 analog mode. *Digital cameras of the higher end variety typically offer 7.5 frames/sec vs 30 (real time) or even less depending on the amount of pixels you need to process. If you wish to do real time imaging 30fps, 7.5fps will not do. Also for fluorescence the shortest duration exposure is preferred. * The camera has to have a good performance to cost ratio. At the low end there are plenty of security type cameras out there, but will they offer you the performance you require. * The big point here is that the higher the signal to noise, the quieter the signal, the more gain that can be added to the signal/ which adds noise as a result, but allows the user to get more sensitivity. Thus if you start with the quietest / highest signal to noise obtainable the better off you are when trying to get more gray levels with minimum sensitivity or more signal to noise allows more gain to be added to achieve a set sensitivity far superior to a signal to noise start point less than the DVC.
The following features listed below are offered by the DVC camera line:
General Statement: Analog video RS-170 focus *Most digitizer boards are typically 8 bits some go higher with digital RS-422 input. With Dc's 10 bits of signal on the analog RS-170 video, this offers first of all more top end room for adding more sensitivity and still maintaining 8 bits/ 256 gray levels for the digitizer board. *You will see more digitizer boards with 10 bit capability on the analog RS-170 input. What will you use as a real time 30 frame / sec camera to match that feature. With most ( real time ) cameras being in the 7 bit range, they would better qualify for security applications. There are some 8 bit cameras, but they only offer 256 gray levels with not much room to add gain for more sensitivity. The DVC camera line offers 10 bits or 1024 gray levels output for those boards along with simultaneous digital RS-422 on the digital models One of the digitizer boards with a 10 bit analog front end is Mutech. They are the only one presently that has that capability for now and others will follow soon.
* Very high signal to noise } 62dB at .5 lux at 30fps/ real time * Frame transfer 1/2" format sensor with 755 horizontal by 484 vertical pixels, or 565 TV lines horizontal by 350 TV lines vertical resolution. * The only upgradable research camera with the researchers funding limitations in mind using the same high quality sensor on all 3 models. Upgradability from the RS-170 analog unit to the dual output 8 bit digital or to the dual output 10 bit digital models with RS-170 analog. (This allows the camera to grow with the researcher) and his budget. See part 3 of 3 next.
And Digitizer Selection Guide For Benefit of Microscope Users Group
Monochrome Digital And Analog Video Camera Line By DVC Company Part 1 of 3 due to capacity limitations of e-mail ***************************************************************** With all due respect: If anyone is offended by a guide to choosing a CCD video camera with a new product release that is applicable for the users in this group ( HIT YOUR DELETE KEY NOW! ) There are some users that might appreciate this information. (This was done with good intentions) so any providers of negative feedback have been given the DELETE KEY OPTION and need not waste net time, thank you. Valid questions are appreciated and I will try to answer them relative to the volume I receive. Included is list of RS-422 digitizer boards in part 2 and 3. * Please address any inquiry directly to dvcco-at-aol.com, not the main file server. ***************************************************************** Dear Microscope Users,
I wish to share some data with you on analog and digital monochrome cameras that I thought you might be able to make use of when reviewing your present or future camera situation. Some of the below information you might be aware of, but others might not. If anyone belongs to other pertinent groups and believes this information to be of benefit to that particular group we would appreciate you letting us know of them.
Monochrome cameras offer much superior gray level and resolution performance along with higher signal to noise specs than the typical color versions. Everyone has different needs and the data below is for researchers who have decided to utilize the benefits of monochrome cameras. Adding pseudo colors to the many more gray levels attainable with a monochrome camera might be a option for you, or adding a tunable liquid crystal filter in line with the camera for serial Red, Green, Blue could also be a option if accurate correlation of the pixel is important. For monochrome applications a electronic tunable filter exists that is very selective with 5,7,10,15,20 ,35 or 50nm band pass filters from 450 to 1050 nm range tunable electronically! See 2 of 2 for info on tunable filters.
Some things to look for in a research grade monochrome camera are the following: * The highest signal to noise possible in a real time 30 frames/sec. unit. Signal to noise is the ultimate measure of a camera and no amount of manipulation, bells and whistles, or 20 different knobs to adjust, will change the bottom line which is the amount of gray levels obtainable. * No geometric distortion- CCD cameras offer basically no distortion while any tube camera gives you distortion in the } 5% range. My opinion is that to use a tube camera where a ccd unit could be substituted is just qualitative, not quantitative microscopy. * No dead pixels on your sensor and lowest pixel variation with no false fill-ins of pixels with data from the pixel next to it by use of memory circuits due to imperfections in the manufacturing process. Different types of CCD cameras have their uses. The 2 main types are frame transfer (FT) and interline transfer (IT) types for microscopy. CID types are also out there but have uses more in machine vision applications. Interline transfer sensors have large gaps between the pixels due to the construction. They offer 100% fill factor by the use of plastic micro lenses that help their sensitivity due to smaller well capacity, but still have the ( same smaller wells.) Plastic does not offer good performance down in the UV range below 400nm, and have other limitations at the near IR range. 1/3 format sensors have even less well capacity and I feel are only produced to provide better yields/profit for the manufacturers for security applications.
Frame transfer sensors which DVC uses have dynamic range of 70dB to start and have no dead pixels and no false fill-ins. The wells are very large and the dark current is {30 electrons at 25C, room temperature. The pixels are contiguous, or within angstroms of electrical separation. Larger wells offer higher signal to noise. Double correlated sample and hold circuits built right into the sensor help reduce noise further on some frame transfer sensors. Frame transfer units offer one half the vertical resolution when doing integration and shuttering due the field transfer technique used. Spectral range with glass face plate attached 400-1100nm. Customers doing (laser) work have reported response in the low 200nm range with face plate removed and up to 1300nm.
Typical bit vs signal to noise ranges: 7 bits = 44dB to technically 50dB but more like 54dB realistically 8 bits = 55dB to 61dB or 256 gray levels 10 bits = 62dB+ for 1024 gray levels. **10 bits offers 4 times the performance of an 8 bit camera. * Another words a camera specification of 50dB falls into the 7 bit range even though manufacturers and sales people might have you believe otherwise. If you have an 8 bit board why not use a 8 bit or higher camera.
Video Camera Selection Guide and DVC Product Release 3 of 3
* Sensitivities in the 1x10-4fc range at over 50% video. When other (IT) type cameras barely see an image these DVC units are offering a perfect image with extra gain to spare and no vertical line noise. Estimate performance at about 5-6 ND filter better sensitivity than 768 type interline transfer sensors. * The only 10 bit digital camera with dual outputs RS-170 & RS-422 and at real time video rates! 30 frames per second * The only camera that has no sensor board. Sensor is physically mounted to the front plate of the camera. Cooling above dew point can then be done to have a truly quantitative camera in that temperature of sensor can be lowered from room temperature or regulated at a constant temperature or our -40C unit can be added. * No need to cool the camera in most applications/ save the money. * The DVC camera line was designed from the ground up to be a digital output camera not a security camera. We use linear well regulated multiple voltage linear power supply and solid C-Mounts with a 360 degree compression ring holding the c-ring firmly in place, not just a little set screw that tends to loosen with time and strip out the threads. * 30dB of additional gain can be added for manual internal adjustable operation accessible via a port from outside the camera, or external gain and offset, or computer controlled gain! 30dB gain vs the others at 20dB because the } 62db performance can fully utilize the 30db range. Where with more than 20db on the IT types you just might end up with snow. * Many other modifications can be done and our units do not come with AGC unless you specify automatics. Automatics are for security cameras. DVC will be responsive to your requirements from the end user to the OEM. * Glass face plate removal to allow the user to easily go down to 300nm or so. Customers with laser applications have reported response in the low 200nm range obviously at low quantum efficiencies.
The DVC camera line is excellent for any high signal to noise or low light application and in real time!
DVC fits a niche between the too pricey and slow non real time digital only video cameras and the security type cameras which don't offer enough of what you need. If you like the resolution DVC can offer our flexibility in being able to upgrade to the digital models is nice to have. If any of you are interested in having a digital camera that works with your Silicon Graphics Indy /Indigo 2 or Sun system, let us know. DVC cameras are made in the USA and have a two year warranty.
* Three models of monochrome camera which are all upgradable to the DVC-10. DVC-0A - RS-170 only with DB-37 connector for remote gain and upgradable to the digital output RS-422 versions offering both outputs. DVC-8 - 8 bit RS-422 digital with RS-170 simultaneously. DVC-10 -10 bit RS-422 digital with RS-170 simultaneously.
*** Below you will find a list of RS-422 digitizer boards that are compatible to the DVC camera line, others are in the works.
****** Compatible Digitizers for RS-170 analog monochrome video are any manufacturer. RS-170 is standard, typically from a BNC connector. Does your board have capability to process RS-170 at 10 bits? The DVC analog output will offer this. Remember if running 10 bit analog RS-170 to a 8 bit board you will only see 8 bits.
Some of these boards have either 8 bit or 10 bit and higher capability. *Compatible digitizers for RS-422 digital video that are plug and play with the DVC digital camera line are listed below:
Manufacturer A-Z Platform Board Name/Model Alacron IBM Alacron Bit Flow IBM/MAC Raptor-VL Coreco IBM F-64, OC-500 Datacube IBM MV-200 Dipix IBM P-360, XPG-1000 Epix IBM 4 Meg, Model 12 EDT SUN S-bus EDT Hyperspeed IBM Hyperspeed Imaging Technology IBM 15040, AFG, MFG Imagraph IBM HI-Def Matrox IBM Magic, 1280, 640, L/C MuTech IBM 10 bit analog video in Precision Digital (PDI) IBM/MAC IMAXX-PCI, SD Univision IBM Piranha
Let DVC know if you wish, the following information: Which digitizer board you are using or would like to use, or need help on. If you have interest in the PCI bus IBM, MAC, Sun, or SGI Indy or Indigo2 computer used If DVC can help answer some of your technical concerns. If your interest is analog or digital or both If your interest is in tunable electronically selectable filters call DVC.
***************************************************************************** ***New DVC Camera Release estimated 4/95 will be the following: Same features as above except now the digital RS-422 can be programmed or reprogrammed for the VINO digital bus on the ((( SGI- Silicon Graphics Indy/Indigo 2 workstations))) Imagine being able to move from SGI to RS-422 digital compatibility with simultaneous RS-170 analog video! Info on our cameras will be on the SGI Web soon. *****************************************************************************
Feel free to e-mail, fax, or call DVC for more information. If DVC can not reach you, we can not help you, so we chose to put this info on the net in hopes of helping anyone that needs it with a cost effective research grade video camera product that we feel, has a definite focus to this microscope group. DVC has heard many of your problems and can be helpful, and possibly offer a solution. Stay in touch.
Sincerely,
Richard Klotsche DVC Company e-mail: dvcco-at-aol.com phone: (619) 444-8300 fax: (619) 444-8321
} I have heard rumors that there will be an international microbeam analysis } meeting in Australia early in 1996. Does anyone know if } this is so; and if it is, do you have any information about when and } where it will be?
The meeting is the first meeting of the International Union of Microbeam Analysis Societies (AKA IUMAS).
It will be held at the University of Sydney, Sydney, NSW Australia for Feb 5 to Feb 9 1996 (Preceeded by workshops) in conjunction with our 14th E.M. Conference and 9th LM symposium. All welcome. Warm weather, water, cool beer. David Cockayne is the chair.
Information on the WWW at http://www.bio.uts.edu.au/em.html
For low cost, high capacity, high resolution and high quality, _nothing_ beats film!
ÒUnfortunatelyÓ if you want to do digital image processing on micrographs then they must be digitised. As previous answers have indicated there is a choice between flat bed scanners and CCD cameras. ThereÕs no doubt that the flatbed scanner produces better data, but the cost is very high. (In addition to purchase price you have maintenance, and so on.). Bill TivolÕs first reply overestimated the cost of the CCD camera solution. You can get away with well under $10000 for camera, lens, stand and light box, a tenth of the price for flat bed scanners.
Optronics made a rotating drum scanner some years ago, but I think itÕs disappeared now. Does anyone know? What did/does it cost?
The Ektron scanner mentioned by Larry Allard is new to me. Does the referenced article give details? Who makes it?
Lab quality CCD cameras often have a geometric distortion between x and y directions of a few percent. This must be measured and corrected for in software. Another limitation is nonlinearity of the response with OD, if you are planning on intensity measurements. Again, it must be measured (with a stepwedge of standard ODs) and corrected for.
CCD cameras have problems with ED patterns, as Bill also pointed out. The very sharp spots with very high gradients cause trouble. I know of some people who are using CCD camera for ED digitising, maybe they can comment in the List about performance.
Another solution is to miss out film and have the CCD camera on the microscope. Horribly expensive, but it has certain advantages. Is anyone digitising ED patterns in this way, and what sort of performance are they getting? I know of one group who is doing so, with success. Are the specifications of on-line CCD cameras so much better than lab ones?
As my old dad used to say ÒYou get what you pay forÓ when confronted with The Sun newspaper at 20p and The Times, at 30p.
I would like to thank everyone that replied to my query concerning networking a Tektronix Phaser IISDX dye sublimation printer. Following is a summary of the problem and the responses.
I wished to put the printer on a Novell network so that it could be accessed from macs, PCs, and Unix boxes. I had purchased Tektronix's Internal Ethertalk card (~$650).
I was told by several listserver users and multiple people at Tektronix that this was absolutely impossible to network the printer under a Novell server using the Ethertalk card. Tektronix put me on to a company in California called Milan that makes a network interface box that should have allowed me to use the parallel port of the printer as the network port. The cost of this box was ~$850.
I was told by several listserver users that it was trivial, no problem, plug it in and it works regarding hooking the printer up to the network.
Finally, I convinced my network administrator to set up a que on one of the network servers and to activate a faceplate so that I could connect my printer to the net. The result .... Novell works fine with Tektronix's Ethertalk card. I have printed from all three platforms and they all work.
The moral of the story: Keep asking your question until you get the answer you want to hear!
p.s. For all of those highly experienced network users who told me in no uncertain terms that I was a fool for not checking with my network administrator before purchasing the printer .... Our local network here has, conservatively speaking, 5,000 users. The network administration is organized with a flowchart of personnel that would make the Ex- Soviet politburo blush. I did try to go through logical channels before making the purchase and was told, "we won't guarantee that it will work, the best we can offer is for you to buy the printer and we'll test it out when it gets here." I suppose this is probably an indication of what the future of networking will be (once everybody's LAN gets so huge).
Sorry for wasting the bandwidth, but last week a 3 part information guide on a particular ccd camera was posted to this forum or the confocal microscopy list server. A disk crash lost my copy of this posting as well as the address of the sender before I had a chance to review it. If the sender could repost a copy directly to me, I would appreciate it. Again, sorry for cluttering your mailbox.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
Well, I was cleaning my email folders on the mainframe and came across this postponed message. Here it is, hope it can still be useful.
Regards, Glen
There is always celloidin, its disadvantages are a lengthy embedding process. It produces a lot of shrinkage in animal tissue, the plant cell wall should resist gross shrinkage, but there may be vacuolization.
For material that has been stained prior to embedding you might try embedding in an epoxy like Pelco's EMBed. You can cut up to 50 microns by either a carbide tungsten knive or using a standard steel knife. These are real knives, not those disposable blades. You also need a solidly built microtome. The carbide knife only needs a slow cutting stroke. The steel knife needs an extremely slow stroke or you can watch the metal forming the knife's edge be ripped away before your very eyes. A small tacking iron, like those used for dry mounting photographs, can be clamped on the end of the knife with a C-clamp. Heat the knife to about 50 deg. C and the plastic will cut more easily.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On Tue, 8 Nov 1994, Mike Folsom wrote:
} Folks - } } A quick question - } } I'm trying to embed plant material is some type of matrix that will } allow me to make fairly thick sections, say 25 - 50 um. } } I've used paraffin and the tissue starts to fracture when section } thickness gets greater than 20 - 25 um. Frankly besides embedding } the tissue in plastic and then grinding it down I'm not sure } what else to do. } } I'd appreciate any suggestions - } } Michael } } _______________________________________________________________________________ } M.W.Folsom/Biology/UNM/Albuquerque,NM~87131/505.277.4277/mwfolsom-at-hydra.unm.edu } }
Message-Id: {199503210352.AA02291-at-shrike.adl.soils.csiro.au} X-Sender: mcc332-at-shrike.adl.soils.csiro.au X-Mailer: Windows Eudora Version 2.0.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Jay Jerome {jjerome-at-isnet.is.wfu.edu}
} Date: Sun, 19 Mar 1995 03:52:28 -0500 } From: DVCCO-at-aol.com } To: Microscopy-at-aaem.amc.anl.gov } Subject: Video Camera Selection Guide1of3 } } Subject: Guide to Choosing A CCD Video Camera With New DVC Product Release } And Digitizer Selection Guide For Benefit of Microscope Users Group } } Monochrome Digital And Analog Video Camera Line By DVC Company } Part 1 of 3 due to capacity limitations of e-mail } ***************************************************************** } With all due respect: If anyone is offended by a guide to choosing a } CCD video camera with a new product release that is } applicable for the users in this group ( HIT YOUR DELETE KEY NOW! ) } There are some users that might appreciate this information. } (This was done with good intentions) so any providers of negative feedback } have been given the DELETE KEY OPTION and need not waste net time, thank you. } Valid questions are appreciated and I will try to answer them relative to } the volume I receive. Included is list of RS-422 digitizer boards in part 2 } and 3. } * Please address any inquiry directly to dvcco-at-aol.com, not the main file } server. } ***************************************************************** } Dear Microscope Users, } } I wish to share some data with you on analog and digital monochrome cameras } that I thought you might be able to make use of when reviewing your present } or future camera situation. Some of the below information you might be aware } of, but others might not. If anyone belongs to other pertinent groups and } believes this information to be of benefit to that particular group we would } appreciate you letting us know of them. } } Monochrome cameras offer much superior gray level and resolution performance } along with higher signal to noise specs than the typical color versions. } Everyone has different needs and the data below is for researchers who have } decided to utilize the benefits of monochrome cameras. Adding pseudo colors } to the many more gray levels attainable with a monochrome camera might be a } option for you, or adding a tunable liquid crystal filter in line with the } camera for serial Red, Green, Blue could also be a option if accurate } correlation of the pixel is important. For monochrome applications a } electronic tunable filter exists that is very selective with 5,7,10,15,20 } ,35 or 50nm band pass filters from 450 to 1050 nm range tunable } electronically! } See 2 of 2 for info on tunable filters. } } Some things to look for in a research grade monochrome camera are the } following: } * The highest signal to noise possible in a real time 30 frames/sec. unit. } Signal to noise is the ultimate measure of a camera and no amount of } manipulation, bells and whistles, or 20 different knobs to adjust, will } change the bottom line which is the amount of gray levels obtainable. } * No geometric distortion- CCD cameras offer basically no distortion while } any tube camera gives you distortion in the } 5% range. My opinion is } that to use a tube camera where a ccd unit could be substituted is just } qualitative, not quantitative microscopy. } * No dead pixels on your sensor and lowest pixel variation with no false } fill-ins of pixels with data from the pixel next to it by use of memory } circuits due to imperfections in the manufacturing process. } Different types of CCD cameras have their uses. } The 2 main types are frame transfer (FT) and interline transfer (IT) } types for microscopy. CID types are also out there but have uses more in } machine vision applications. } Interline transfer sensors have large gaps between the pixels due to the } construction. They offer 100% fill factor by the use of plastic } micro lenses that help their sensitivity due to smaller well capacity, } but still have the ( same smaller wells.) Plastic does not offer good } performance down in the UV range below 400nm, and have other limitations } at the near IR range. 1/3 format sensors have even less well capacity } and I feel are only produced to provide better yields/profit for the } manufacturers for security applications. } } Frame transfer sensors which DVC uses have dynamic range of 70dB to start } and have no dead pixels and no false fill-ins. The wells are very } large and the dark current is {30 electrons at 25C, room temperature. } The pixels are contiguous, or within angstroms of electrical separation. } Larger wells offer higher signal to noise. Double correlated sample and } hold circuits built right into the sensor help reduce noise further on } some frame transfer sensors. Frame transfer units offer one half the } vertical } resolution when doing integration and shuttering due the field transfer } technique used. } Spectral range with glass face plate attached 400-1100nm. Customers } doing (laser) work have reported response in the low 200nm range with } face plate removed and up to 1300nm. } } Typical bit vs signal to noise ranges: } 7 bits = 44dB to technically 50dB but more like 54dB realistically } 8 bits = 55dB to 61dB or 256 gray levels } 10 bits = 62dB+ for 1024 gray levels. **10 bits offers 4 times the } performance of an 8 bit camera. } * Another words a camera specification of 50dB falls into the 7 bit range } even though manufacturers and sales people might have you believe } otherwise. If you have an 8 bit board why not use a 8 bit or higher } camera. } } } --------------------------------------------------------------------------- Stuart G. McClure, | Post small : P.B.#2, Glen Osmond, CSIRO Division of Soils, | Sth Australia, AUSTRALIA, 5064. Adelaide Laboratories, | Post large : Waite Rd, Urrbrae, SA, Australia | Sth Australia, AUSTRALIA, 5064.
Phone: (08) 303-8484 International use +61-8- instead of (08) Fax: (08) 303-8550 Email: Stuart.McClure-at-adl.soils.csiro.au ---------------------------------------------------------------------------
Message-Id: {199503210352.AA02295-at-shrike.adl.soils.csiro.au} X-Sender: mcc332-at-shrike.adl.soils.csiro.au X-Mailer: Windows Eudora Version 2.0.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Jay Jerome {jjerome-at-isnet.is.wfu.edu}
} Date: Sun, 19 Mar 1995 03:54:20 -0500 } From: DVCCO-at-aol.com } To: Microscopy-at-aaem.amc.anl.gov } Subject: Video Camera Selection Guide2of3 } } DVC Video Camera Guide And Release 2 of 2 } } Remember, if the camera is on the ragged edge of 8 bit performance and } you want to add gain for more sensitivity, even the smallest amount of } gain might put you in the 7 bit range if you we not there already. } If you have 10 bit capability on your board why not use a 10 bit camera. } * Also a camera specification of 60dB is not a 10 bit camera only an 8. } * Also there are many types of noise in a sensor besides thermal noise. } Cooling a sensor will reduce (thermal noise only) and allow you to } integrate longer. I would double check the impression some make that } when a 50db sensor is cooled it offers a better s/n of 10 or 12 bits vs } just 7. } The reduction of one type of noise (thermal) is all that cooling } provides. } * Other types of sensor noise besides dark current include photon, nyquest, } shot, gain noise, and flicker noise etc. } * The signal to noise of the sensor at room temperature is a good measure } of what you can expect. } ** Look at the quantum efficiency at the nm range needed. } ** look at the spectral response of the sensor. } ** There are different grades of sensors available relative to dead } pixels and gray level variation on other manufacturers cameras. } *Digital RS-422 eliminates pixel jitter for sub-pixel accuracy if it is } needs versus the genlock action between the digitizer board and the } camera as they try to lock to each other in the RS-170 analog mode. } *Digital cameras of the higher end variety typically offer 7.5 frames/sec } vs 30 (real time) or even less depending on the amount of pixels you need } to process. If you wish to do real time imaging 30fps, 7.5fps will not } do. Also for fluorescence the shortest duration exposure is preferred. } * The camera has to have a good performance to cost ratio. At the low end } there are plenty of security type cameras out there, but will they offer } you the performance you require. } * The big point here is that the higher the signal to noise, the quieter } the signal, the more gain that can be added to the signal/ which adds noise } as a result, but allows the user to get more sensitivity. } Thus if you start with the quietest / highest signal to noise obtainable } the better off you are when trying to get more gray levels with minimum } sensitivity or more signal to noise allows more gain to be added to achieve } a set sensitivity far superior to a signal to noise start point less than } the DVC. } } The following features listed below are offered by the DVC camera line: } } General Statement: Analog video RS-170 focus } *Most digitizer boards are typically 8 bits some go higher with digital } RS-422 } input. With Dc's 10 bits of signal on the analog RS-170 video, this offers } first of all more top end room for adding more sensitivity and still } maintaining } 8 bits/ 256 gray levels for the digitizer board. } *You will see more digitizer boards with 10 bit capability on the analog } RS-170 } input. What will you use as a real time 30 frame / sec camera to match that } feature. With most ( real time ) cameras being in the 7 bit range, they } would } better qualify for security applications. There are some 8 bit cameras, but } they only offer 256 gray levels with not much room to add gain for more } sensitivity. The DVC camera line offers 10 bits or 1024 gray levels output } for those boards along with simultaneous digital RS-422 on the digital models } One of the digitizer boards with a 10 bit analog front end is Mutech. They } are the only one presently that has that capability for now and others will } follow soon. } } * Very high signal to noise } 62dB at .5 lux at 30fps/ real time } * Frame transfer 1/2" format sensor with 755 horizontal by 484 vertical } pixels, or 565 TV lines horizontal by 350 TV lines vertical resolution. } * The only upgradable research camera with the researchers funding } limitations in mind using the same high quality sensor on all 3 models. } Upgradability from the RS-170 analog unit to the dual output 8 bit } digital or to the dual output 10 bit digital models with RS-170 analog. } (This allows the camera to grow with the researcher) and his budget. } See part 3 of 3 next. } } --------------------------------------------------------------------------- Stuart G. McClure, | Post small : P.B.#2, Glen Osmond, CSIRO Division of Soils, | Sth Australia, AUSTRALIA, 5064. Adelaide Laboratories, | Post large : Waite Rd, Urrbrae, SA, Australia | Sth Australia, AUSTRALIA, 5064.
Phone: (08) 303-8484 International use +61-8- instead of (08) Fax: (08) 303-8550 Email: Stuart.McClure-at-adl.soils.csiro.au ---------------------------------------------------------------------------
Message-Id: {199503210352.AA02299-at-shrike.adl.soils.csiro.au} X-Sender: mcc332-at-shrike.adl.soils.csiro.au X-Mailer: Windows Eudora Version 2.0.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Jay Jerome {jjerome-at-isnet.is.wfu.edu}
} Date: Sun, 19 Mar 1995 03:54:55 -0500 } From: DVCCO-at-aol.com } To: Microscopy-at-aaem.amc.anl.gov } Subject: Video Camera Selection Guide3of3 } } Video Camera Selection Guide and DVC Product Release 3 of 3 } } * Sensitivities in the 1x10-4fc range at over 50% video. When other (IT) } type cameras barely see an image these DVC units are offering a perfect } image with extra gain to spare and no vertical line noise. Estimate } performance at about 5-6 ND filter better sensitivity than 768 type } interline transfer sensors. } * The only 10 bit digital camera with dual outputs RS-170 & RS-422 and at } real time video rates! 30 frames per second } * The only camera that has no sensor board. Sensor is physically mounted } to the front plate of the camera. Cooling above dew point can then be } done to have a truly quantitative camera in that temperature of sensor } can be lowered from room temperature or regulated at a constant } temperature or our -40C unit can be added. } * No need to cool the camera in most applications/ save the money. } * The DVC camera line was designed from the ground up to be a digital } output camera not a security camera. We use linear well regulated } multiple voltage linear power supply and solid C-Mounts with a 360 } degree compression ring holding the c-ring firmly in place, not just a } little set screw that tends to loosen with time and strip out the } threads. } * 30dB of additional gain can be added for manual internal adjustable } operation accessible via a port from outside the camera, or external gain } and offset, or computer controlled gain! 30dB gain vs the others at 20dB } because the } 62db performance can fully utilize the 30db range. Where } with more than 20db on the IT types you just might end up with snow. } * Many other modifications can be done and our units do not come with AGC } unless you specify automatics. Automatics are for security cameras. } DVC will be responsive to your requirements from the end user to the OEM. } * Glass face plate removal to allow the user to easily go down to 300nm or } so. Customers with laser applications have reported response in the low } 200nm range obviously at low quantum efficiencies. } } The DVC camera line is excellent for any high signal to noise or low light } application and in real time! } } DVC fits a niche between the too pricey and slow non real time digital only } video cameras and the security type cameras which don't offer enough of what } you need. If you like the resolution DVC can offer our flexibility in being } able to upgrade to the digital models is nice to have. } If any of you are interested in having a digital camera that works with your } Silicon Graphics Indy /Indigo 2 or Sun system, let us know. } DVC cameras are made in the USA and have a two year warranty. } } * Three models of monochrome camera which are all upgradable to the DVC-10. } DVC-0A - RS-170 only with DB-37 connector for remote gain and upgradable } to the digital output RS-422 versions offering both } outputs. } DVC-8 - 8 bit RS-422 digital with RS-170 simultaneously. } DVC-10 -10 bit RS-422 digital with RS-170 simultaneously. } } *** Below you will find a list of RS-422 digitizer boards that are compatible } to the DVC camera line, others are in the works. } } ****** Compatible Digitizers for RS-170 analog monochrome video are any } manufacturer. RS-170 is standard, typically from a BNC connector. } Does your board have capability to process RS-170 at 10 bits? } The DVC analog output will offer this. Remember if running 10 bit analog } RS-170 to a 8 bit board you will only see 8 bits. } } Some of these boards have either 8 bit or 10 bit and higher capability. } *Compatible digitizers for RS-422 digital video that are } plug and play with the DVC digital camera line are listed below: } } Manufacturer A-Z Platform Board Name/Model } Alacron IBM Alacron } Bit Flow IBM/MAC Raptor-VL } Coreco IBM F-64, OC-500 } Datacube IBM MV-200 } Dipix IBM P-360, XPG-1000 } Epix IBM 4 Meg, Model 12 } EDT SUN S-bus EDT } Hyperspeed IBM Hyperspeed } Imaging Technology IBM 15040, AFG, MFG } Imagraph IBM HI-Def } Matrox IBM Magic, 1280, 640, L/C } MuTech IBM 10 bit analog video in } Precision Digital (PDI) IBM/MAC IMAXX-PCI, SD } Univision IBM Piranha } } Let DVC know if you wish, the following information: } Which digitizer board you are using or would like to use, or need help on. } If you have interest in the PCI bus } IBM, MAC, Sun, or SGI Indy or Indigo2 computer used } If DVC can help answer some of your technical concerns. } If your interest is analog or digital or both } If your interest is in tunable electronically selectable filters call DVC. } } ***************************************************************************** } ***New DVC Camera Release estimated 4/95 will be the following: } Same features as above except now the digital RS-422 can be programmed or } reprogrammed for the VINO digital bus on the } ((( SGI- Silicon Graphics Indy/Indigo 2 workstations))) } Imagine being able to move from SGI to RS-422 digital compatibility } with simultaneous RS-170 analog video! } Info on our cameras will be on the SGI Web soon. } ***************************************************************************** } } Feel free to e-mail, fax, or call DVC for more information. } If DVC can not reach you, we can not help you, so we chose to put this info } on } the net in hopes of helping anyone that needs it with a cost effective } research } grade video camera product that we feel, has a definite focus to this } microscope group. DVC has heard many of your problems and can be helpful, } and } possibly offer a solution. Stay in touch. } } Sincerely, } } Richard Klotsche } DVC Company } e-mail: dvcco-at-aol.com } phone: (619) 444-8300 } fax: (619) 444-8321 } } } } } } } --------------------------------------------------------------------------- Stuart G. McClure, | Post small : P.B.#2, Glen Osmond, CSIRO Division of Soils, | Sth Australia, AUSTRALIA, 5064. Adelaide Laboratories, | Post large : Waite Rd, Urrbrae, SA, Australia | Sth Australia, AUSTRALIA, 5064.
Phone: (08) 303-8484 International use +61-8- instead of (08) Fax: (08) 303-8550 Email: Stuart.McClure-at-adl.soils.csiro.au ---------------------------------------------------------------------------
With reference to embedding plant material for thick(ish) sections. We routinely use LR White (London Resin White acrylic resin for plant material and can certainly cut sections up to 15-20um. The nice thing about LR white is that is like the British, tolerant and kind to specimens and you need only dehydrate to 80% EtOH and still get godd embedding. You can use all sorts of light microscopy stains on sections cut from such material.
Greetings from a beautiful spring morning in Cambridge where all the daffodils are in full bloom along the Backs.
Patrick Echlin
Multi-Imaging Centre School of Biological Sciences
On Mon, 20 Mar 1995, Glen Macdonald wrote:
} Well, I was cleaning my email folders on the mainframe and came across } this postponed message. Here it is, hope it can still be useful. } } Regards, } Glen } } } There is always celloidin, its disadvantages are a lengthy embedding } process. It produces a lot of shrinkage in animal tissue, the plant } cell wall should resist gross shrinkage, but there may be vacuolization. } } For material that has been stained prior to embedding you might try } embedding in an epoxy like Pelco's EMBed. You can cut up to 50 microns } by either a carbide tungsten knive or using a standard steel knife. } These are real knives, not those disposable blades. You also need a } solidly built microtome. } The carbide knife only needs a slow cutting stroke. The steel knife } needs an extremely slow stroke or you can watch the metal forming the } knife's edge be ripped away before your very eyes. A small tacking iron, } like those used for dry mounting photographs, can be clamped on the end of } the knife with a C-clamp. Heat the knife to about 50 deg. C and the } plastic will cut more easily. } } Glen MacDonald } Hearing Development Laboratories RL-30 } University of Washington } Seattle, WA 98195 } (206)543-8360 } glenmac-at-u.washington.edu } } } On Tue, 8 Nov 1994, Mike Folsom wrote: } } } Folks - } } } } A quick question - } } } } I'm trying to embed plant material is some type of matrix that will } } allow me to make fairly thick sections, say 25 - 50 um. } } } } I've used paraffin and the tissue starts to fracture when section } } thickness gets greater than 20 - 25 um. Frankly besides embedding } } the tissue in plastic and then grinding it down I'm not sure } } what else to do. } } } } I'd appreciate any suggestions - } } } } Michael } } } } _______________________________________________________________________________ } } M.W.Folsom/Biology/UNM/Albuquerque,NM~87131/505.277.4277/mwfolsom-at-hydra.unm.edu } } } } } }
I'm a recent subscriber and am wondering if perhaps there is another listserver that might be closer to my fields of interest, i.e. micro-manipulation of cellular and sub-cellular particles either via mechanica l or optical trapping techniques. If anyone knows of a group that is more involved in these areas, I would appreciate if you could reply to me directly.
WINTHROP UNIVERSITY Electronic Mail Message Date: 21-Mar-1995 12:50pm EST From: Kenneth Gregg GREGGK Dept: Biology Tel No: 803-323-2111
TO: Remote Addressee ( _smtp%"microscopy-at-aaem.amc.anl.gov" )
WINTHROP UNIVERSITY Electronic Mail Message Date: 21-Mar-1995 11:31am EST From: Kenneth Gregg GREGGK Dept: Biology Tel No: 803-323-2111
TO: Remote Addressee ( smtp%"microscopy-at-aaem.amc.anl.gov" )
Greetings, My guess is that many members of the list teach a histology course at the University level. I would like some comments on a method of teaching the laboratory section of histology which we are considering. We find that many students are rapidly bored when looking at tissue/organ sections in the light microscope. They also seem to lack the ability to know when they have acquired an acceptable level of expertise. As an antidote to this we are kicking around the following idea -- and we would like to know if any members of the list have tried similar approaches, and with what success?
We are thinking about having individual students prepare computer-assisted-instruction lessons on topics such as epithelial tissues, liver structure, etc. We would like to have the students prepare a hypertext program for each topic. They would integrate the hypertext with captured images from their light microscope observations as well as scanned diagrams and electron micrographs which they could obtain from texts or photographs. Students would be very much engaged in their own productions and could study lessons prepared by other students. These programs could be used and improved in subsequent years by other students. They could also be made available for other institutions.
If you prefer, you may respond to me personally at Greggk-at-Winthrop.edu
We are seriously interested in purchasing a setup for our Nikon Diaphot that could be used for the following: o quantitation of very low level fluorescence-- sensitive and linear o but also useful for critical Nomarski applications such as microtubule motility assays o automation of filter wheels for multiple probes or fluorescence / phase contrast image collection o automation of stage moving, for instance for making mosaics automatically or being able to move back to a specific field o maybe Z motor and deconvolution, although the are not our main concerns o ESSENTIAL: we are a multi-user facility so we need ease of operation
We are considering a Photometrics or Princeton Instruments cooled CCD camera. We have looked at Oncor Imaging, I.P. Lab and have spoken to other companies. For hardware, we have called companies that offer individual parts (e.g. just the filter wheels) and, of course, have checked out Ludl. We love NIH-Image, but want something does not require a large amount of macro or other programming to get the hardware to work; basically, we want plug and play. Any suggestions or "stay away from this product at all costs" comments on parts of systems or whole systems would be appreciated. Thanks- Michael Cammer cammer-at-aecom.yu.edu
Hi there, Could you help me with the problem of delineation of titanium in bone and soft tissue.Any information and/or references would be a great help to me. Study will involve the use of both SEM and EPMA, possibly FTIR as well. Thank you all, Wis Jablonski , Email W.Jablonski-at-csl.utas.edu.au
Place: Convene at Forensic Science Associates in Richmond, Conclude at Oakland PD. (See contact information below for reservations, maps, car pooling, and further details.)
Time: Convene by 10:00 am, Adjourn by 5:00 pm
Summary: This promises to be a most informative and interesting meeting as we hear from some of the Bay Areas foremost forensic microscopists, see some of the latest technology in microscopy, and hear about some interesting forensic applications of microscopy.
Agenda:
Forensic Science Associates, 10:00 am. Stephen A. Shaffer of MicroDataware will introduce the topic of forensic science in general and forensic microscopy in particular. Peter D. Barnett of Forensic Science Associates will then discuss two interesting cases involving forensic microscopy, and also the role of the consulting (i.e., non-government) microscopist in forensic cases. Pete will discuss cases involving questions of ballot marking and alleged surgical mistakes revealed by fibers in the nose! We'll conclude at FSA by 11:30 or 11:45.
Bureau of Alcohol, Tobacco, and Firearms, 1:00 pm. After a brief break for lunch at a Deli in Walnut Creek, we'll re-convene at the ATF Laboratory where we will divide into two groups to cover separate topics with two speakers. John Murdock will explain and demonstrate the use of comparison microscopy in firearms identification. John is the former Director of the Contra Costa County Crime Laboratory and also teaches criminalistics locally. He is an excellent speaker and promises a fascinating discussion. Also at ATF, we will meet with Robert Thompson who will demonstrate a new technology for automated comparison microscopy. This technology may revolutionize firearms work in criminalistics laboratories by screening cases and suggesting most promising comparisons for the human microscopist to perform. We will adjourn from ATF by 2:30 to proceed to the Oakland Police Department.
Oakland Police Department, 3:00 pm. At OPD, Diane Bowman will discuss the use of microchemical testing procedures for the identification of illicit drugs and narcotics, showing both the utility and the beauty of these procedures for the rapid identification of controlled substances. Mary Gibbons, Director of the OPD Lab, will discuss a fascinating case where microscopic particles of spray paint, identified and compared by microscopy, were instrumental in unraveling a suspect's story, leading to solution of a homicide. We will aduourn from OPD by 5:00 pm.
At both ATF and OPD, each of our sub-groups will take turns with each of the speakers so we all get to hear all of the topics. Each topic will be covered in a presentation of approximately 45 minutes duration.
Reservations Required!!! Because of the limited space available within the microscopy labs we will visit, it may be necessary to limit the number of people who attend. No more than 20 can be accommodated so call Steve Shaffer ASAP to reserve your space, receive maps to the locations, and to coordinate with others on car pooling to the three sites we will visit.
Contact: Stephen A. Shaffer at the numbers below. Don't delay, call right away!
************************************************************ * Stephen A. Shaffer * Publishers of The Particle Atlas * * MicroDataware * on CD-ROM. Developers of custom * * 2894 Tribune Avenue * image and database software for * * Hayward CA 94542-1637 * laboratories, specializing in * * 1-510-582-6624 voice * light and electron microscopy. * * 1-510-582-6624 fax * Email inquiries invited. * * sshaffer-at-microdataware.com * ************************************************************
Plant material can also be embedded in butyl methyl methacrylate and sectioned up to 20 um. Stick the sections down to glass slides with polyethylenimine. The resin is removed with acetone before antibody staining, but you don't need to remove it if staining with something small and water-soluble like aniline blue.
____________________________________________________________ Rosemary White __ / Department of Ecology _/ \__/ \ and Evolutionary Biology / \ Monash University / Australia \ Clayton, Victoria 3168 \ ____ / phone 61-3-905 5670 \_/ \_*_/ fax 61-3-905 5613 __ email r.g.white-at-sci.monash.edu.au \/
Message-Id: {1995Mar21.151709.2878643829-at-dmcmail.ucsf.edu} To: microscopy-at-aaem.amc.anl.gov (microscopy)
Subject: Time: 2:10 PM OFFICE MEMO Usage "Tracking" Programs Date: 3/21/95 Hi everybody! Does anyone have any usage tracking programs, or know where they might be obtained? I am especially interested in a DOS program that is compatible with Meridian software. Does anyone have any experience using tracking programs? I am also interested in programs that are compatible with HP and Macintosh acquisition/analysis software for flow cytometers (primarily Becton Dickinson.) Any suggestions or comments would be greatly appreciated. Thank you very much!
__________________________________________________________ Kris Kavanau Internet: kavanau-at-dmc.ucsf.edu Manager, Lab for Cell Analysis Telephone: 415-476-2631 Division of Molecular Cytometry FAX: 415-476-8218 University of California San Francisco, CA 94143 __________________________________________________________
-- [ From: Charles A. Garber, PH. D. * EMC.Ver #2.10P ] --
SPI Supplies
For TEM examination, we using the following procedure:
1] Pick up the unstained sections on a silicon dioxide coated TEM grid,
2] Gently etch (e.g. 10 seconds) the now supported section in a barrel geometry reactive etcher (for example, the SPI Model Plasma Prep II) using oxygen as the etching gas, which will immediately etch away all traces of the organic matrix, leaving the inorganics dispersed, in their original locations, on the silicon dioxide support film. Usually these residues form a ghost type of outline of original cell shapes. I don't know what bone might look like if etched this way.
The reason for the silicon dioxide support film is that it will not be etched by the oxygen plasma, where as any kind of organic or carbonaceous support film would otherwise be etched away.
The reason for the etching to remove the organic part of the sample is to reduce to almost zero the Bremstrahlung radiation, thereby increasing greatly the lower detection limits.
3] We have always found the TEM to be better for doing this kind of analysis than SEM/EDS, mainly because of the higher resolution possibilities.
Let me know if you would need further information. The making of the silicon dioxide coated grids requires a bit of art but they are not all that difficult to make, so long as you are patient.
Charles A. Garber, Ph. D. PRESIDENT SPI SUPPLIES PO Box 656 West Chester, PA 19381-0656 USA
The following position is available at the Johns Hopkins U. School of Medicine:
Electron Micrscopy Technician/19 hrs/wk PART-TIME:
Position will be responsible for semi- and ultra-thin sectioning on glass and diamond knives. Standard grid prep and staining of sections; maintenance of electron gun parts (polishing cleaning etc.). Replenising of photo- and lab chemical stocks, care of peripheral equipment and other related duties. College degree, BS or equivalent and minimum two years experience. Special skills/knowledge of diamond knive use and maintenance, routine EM lab operations.
All interested candidates should contact: Lou Cote (410) 955-0519 Job # M.3531.94
Kris et al, I use DIRECT ACCESS from Fifth Generation Systems. It keeps track of the time, reqires a pssword and has the option of also requiring a project name. This forces the issue with people who think it's better for me to chase down their grant number then for them to remember, and makes billing much easier. We also keep a log book for people to record problems and i modify billings accordingly. When I came here this was already installed on the XRD computer so I did no comparative shopping, & I don't know what else is out there. Their address is FIFTH GENERATION SYSTEMS INC 10049 n. Reiger Rd Baton Rouge LA 70809 (504) 291-7221
Maggy Piranian Memorial U of Newfoundland
On Tue, 21 Mar 1995 Kris_Kavanau-at-dmcmail.ucsf.edu wrote:
} Subject: Time: 2:10 PM } OFFICE MEMO Usage "Tracking" Programs Date: 3/21/95 } Hi everybody! } Does anyone have any usage tracking programs, or know where they might be } obtained? I am especially interested in a DOS program that is compatible } with Meridian software. Does anyone have any experience using tracking } programs? I am also interested in programs that are compatible with HP } and Macintosh acquisition/analysis software for flow cytometers (primarily } Becton Dickinson.) } Any suggestions or comments would be greatly appreciated. Thank you very } much! } } __________________________________________________________ } Kris Kavanau Internet: } kavanau-at-dmc.ucsf.edu } Manager, Lab for Cell Analysis Telephone: 415-476-2631 } Division of Molecular Cytometry FAX: 415-476-8218 } University of California } San Francisco, CA 94143 } __________________________________________________________ } } } } }
We regrettably learned Monday, of the untimely death this last Saturday of Mort Maser an individual whose efforts and contributions to the Microscopy Community here in the US are well known and valued by many of us.
Mort will be certainly missed and may I speak for the entire MSA family in expressing our condolences to his family and friends. For those of you who are interested I have gotten some details concerning a memorial for Mort and detail those below.
In memory of Mort, his family is establishing the Morton D Maser Scholarship Fund. Contributions can be sent to: Larry Maser, P.O. Box EM, Woods Hole, MA 02543. Please designate your gift to the Morton D Maser Scholarship Fund.
Memorial services celebrating Mort's life are being planned for the summer months. Details will be announced as they are finalized.
Morton D "Mort" Maser, President of Woods Hole Educational Associates, and Executive Secretary to Council of both the Microscopy Society of America and the Histochemical Society, died on Saturday, March 18, 1995 at the age of 60.
He earned his A.B. from the University of Pennsylvania in 1955, and his Ph.D. in biophysics from the University of Pittsburgh in 1962. He held research and faculty positions at the Mellon Institute, Harvard University, the Millard Fillmore Hospital, Northeastern University, Creighton University, and the Marine Biological Laboratory.
He developed roughly 100 short courses in electron microscopy and related fields for scientists and technicians, and made more than 50 contributions to peer-reviewed publications on electron microsopy and related topics.
He was a member of several professional societies including serving as Chair of EMSA's Education Committee; past President, Director of the New England Society for Electron Microscopy; the American Association for the Advancement of Science; American Society of Cell Biology; and the International Society for Stereology.
His research has spanned the sciences from electron microscopical techniques, to computer sciences, to software systems.
He is survived by his wife, Naomi, children Larry of Forestdale, Massachusetts and Jill, of Philadelphia, and grandson, Benjamin.
I was following the discussion on flatbed scanners with great interest, but I need to find some vendors to request additional information. The following units were mentioned: Umax PowerLink AGFA Arcus Perkin-Elmer and Optronics
Does any one have address, phone number or E-mail address for these or other vendors?
In message Mon, 20 Mar 95 14:23:11 EST, forbes-at-cip.org.ec writes: We would greatly appreciate } suggestions on other ways to clear and fix this tissue that do not } involve lactophenol or other products which require special } disposal facilities, which currently do not exist in Quito. *******
We routinely use 1 M NaOH to clear flowers to check in vivo pollen growth after Aniline Blue staining (for fluorescence). You might also want to try a clearing protocol specifically designed for fungal infected leaf tissue such as the one in the following publication:
AU HIGNIGHT-K-W. MUILENBURG-G-A. VAN-WIJK-A-J-P.
TI A CLEARING TECHNIQUE FOR DETECTING THE FUNGAL ENDOPHYTE ACREMONIUM- SP
IN GRASSES
SO BIOTECH HISTOCHEM
68 (2). 1993. 87-90.
JT BIOTECHNIC & HISTOCHEMISTRY.
KW FESTUCA-ARUNDINACEA FESTUCA-OVINA LOLIUM-PERENNE BRIGHT FIELD
MICROSCOPY ANALYTICAL METHOD
AB Leaf tissue of tall fescue Festuca arundinacea Schreb., hard fescue
Festuca ovina L., red fescue Festuca rubra L. and Perennial ryegrass
Lolium perenne L. was stained with rose Bengal or aniline blue to
detect the presence of the fungal endophyte Acremonium sp.. Specimens
were cleared using methyl salicylate, an optical clearing agent, and
viewed using bright field microscopy. Tissue was preserved as dried
tissue or stored in 70% aqueous ethyl alcohol before staining and
clearing. Tissue was observed at 2, 4 and 12 weeks following clearing
to check for stain retention. Staining with rose Bengal was inferior to
aniline blue when followed by the clearing agent methyl salicylate.
Fungal mycelia stained lighter with rose Bengal and were more difficult
to detect than mycelia stained with aniline blue. The results
illustrate the usefulness of combining staining and methyl salicylate
clearing for detecting fungal endophytes.
___________
Good Luck!
************************************************************************* M.V. Parthasarathy Professor of Plant Biology Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14850 Telephone: (607) 255-1734 Fax: (607) 255-5407 E-mail: mvp2-at-cornell.edu ***********************************************************************
Have you tried calling Universal Imaging (you mentioned some of the other competitive companies). They are at 610-344-9410. Their new metamorph system will do just about everything you requested...and works well with a cooled CCD. Nina Allen
} Date sent: Mon, 27 Feb 1995 09:30:43 -0500 (EST) } From: YANGA-at-NCCCOT.AGR.CA } Subject: RE:35mm film in TEM } To: microscopy-at-aaem.amc.anl.gov
} We use Eastman motion picture film (5302 fine grain release } positive film) in Philips EM300 in the past and currently in } Zeiss EM902 without any problem. } } Ann Fook Yang
We use copex PET10 in our CM12 and 301. It doesn't outgase much, is very thin so is easy to handle and has not sprocket hole to interfere with the images (depending onthe format).
I was following the discussion on flatbed scanners with great interest, but I need to find some vendors to request additional information. The following units were mentioned: Umax PowerLink AGFA Arcus Perkin-Elmer and Optronics
Does any one have address, phone number or E-mail address for these or other vendors?
} To: microscopy-at-aaem.amc.anl.gov } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } Subject: EM TECH POSITION } } The following position is available at the Johns Hopkins U. School of Medicine: } } Electron Micrscopy Technician/19 hrs/wk PART-TIME: } } Position will be responsible for semi- and ultra-thin sectioning on } glass and diamond knives. Standard grid prep and staining of sections; maintenance } of electron gun parts (polishing cleaning etc.). Replenising of photo- and lab } chemical stocks, care of peripheral equipment and other related duties. } College degree, BS or equivalent and minimum two years experience. Special } skills/knowledge of diamond knive use and maintenance, routine EM lab operations. } } All interested candidates should contact: } Lou Cote } (410) 955-0519 } Job # M.3531.94 } } or } } Mike Delannoy } (410) 955-1365 }
Dear Friends, As you all may remember, I was having crossing over problem between FITC & Rhodamine. I followed the suggestion that some of you gave me and used Cy5 (1.4 mg/ml) 1:100 and 1:400 and Cy3 (1.4 mg/ml) same dilution. I did a control and I got a lot background staining. Any suggestion? My next step will be to check my filters as some of you suggested. Nichole Dutton suggested that before applying primaries antibodies to block for nonspecifics with 20% horse serum solution, and use 1% bovine serum albumin in my diluents. Nichole, I'd like further information on this. Dave, also suggested using different blocking reagent (BSA, milk, gelatin, serum.) I'd like to learn more about this possibility, Dave. Thanks, Ciprian
__________________________________________________________ Ciprian Almonte Medical College of Pennsylvania Dept. of Anatomy and Neurobiology E-mail: almonte-at-medcolpa.edu 3200 Henry Ave. Voice: (215) 842-4081 Philadelphia, PA 19129 Fax: (215) 843-9082 __________________________________________________________
Dear Frank, There was a used Perkin-Elmer scanner available, which I heard about from the microscopy list. I don't remember the company name, since we didn't have the money to get the scanner. They sent me a write-up with specs, but I don't know whether I still have it. Perhaps you or Nestor can search the ar- chives for the posting. Good luck. Yours, Bill Tivol
REGARDING Position Available for FIB Engineer Position available for a FIB engineer to join a team working on process development and failure analysis of ULSI circuits in Motorola, Advanced Products Research and Development Laboratory, Austin, Texas. The engineer will be responsible for the setup and operation of a focused-ion-beam facility. The candidate must have hands-on experience on FIB. Qualified candidates please send resumes to
ra2169-at-email.mot.com
or mail to
David Hwang Motorola APRDL, K-10 3501 Ed Bluestein Boulevard Austin, TX 78721
MR-Received: by mta GATEV3; Relayed; Thu, 23 Mar 1995 12:19:38 -0600 (CST) Alternate-recipient: prohibited Disclose-recipients: prohibited
Hi, I am a TEM sample prep technician and is currently preparing samples by self supportive dimpling, tripod and acid etch(mainly for planview)techniques. I am interested in new methods out there being developed and used successfully. Is anyone out there preparing samples using FIB(Focus Ion Beam) as a tool.We have used it once in a while to thin our samples prepared with the wedge technique in the past, but would like to know more.We own a FEI800, but is currently used extensively for SEMs. Also any tips,and/or problems to avoid, associated with the sample prep(especially in the milling dept.) are most welcome. Most of my contamination problem occurs in the ion-milling process. Thanks, Lata Prabhu
Hello all- I am looking for recommendations for evaporation material used in rotary shadowing of proteins. I am referencing the text "Electron Microscopy in Molecular Biology", a practical approach; ed. by Sommerville and Scheer. The method described uses a Balzers electron beam gun to evaporate tungsten/tantalum. We do not have such a gun but plan to use a standard thermal evaporation. Anyone out there have experience/recommendations for evaporation metal, e.g., platinum, tungsten, palladium, carbon/platinum for finer grains?
Thanks, Doug Davis EML Berkeley (510) 642-2085 doug_davis-at-maillink.berkeley.edu
Job opening: Staff Research Associate II Electron Microscope Laboratory University of California at Berkeley $29,200 per annum, 80-100% appointment Job listing #03-345-30/SL closing date: 4-14-95 Contact the Office of Employment Room 7-G 2200 University Ave., Berkeley, CA 94720 (510) 642-1011 general info
Operate and maintain an SEM. Train users in SEM technique, including specimen preparation. Learn operation of JEOL 9000 freeze-fracture machine and train and supervise others in its use. Prepare samples for immunoelectron microscopy (TEM) and occasionally supervise TEM users. Operate cryofixation equipment for EM sample preparation . Train users in darkrom technique and supervise use of lab computers. Qualifications: Experience in cryofixation and immunocytochemical methods. General laboratory skills (preparation of buffer solutions, operating pH meters etc.). Familiarity with using computer programs (word processing, spreadsheets). Familiarity with electronic equipment and its routine maintenance. Darkroom experience. Ability to work independently.
Personnel will not send out job applications. If the applicants cannot come to the personnel office to pick up an application they can send a:
1. Cover Letter referencing the job number (03-345-30) 2. Resume
The cover letter and resume should be mailed to:
University of California Berkeley Berkeley Campus Employment Office Room 7-G (Ground Floor) 2200 University Ave. Berkeley, Ca. 94720
A message forwarded from my colleague Paul Sutherland
Has anyone got a good fixation method for insect gut tissue which has a thick peritrophic membrane. At present I am using 2% paraformaldehyde, 2.5% glutaraldehyde in 0.1M phosphate buffer pH 7.2 and a post fix in 1% osmium tetroxide. With this fix the microvilli are not well preserved.
Ian Hallett
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660
We are trying to localise enzyme sites (ATP) on the plasmalemma of salt glands in halophytic grasses. Has anyone out there got a method(s) for ATP-ase at the LM level for glut. fixed tissue already embedded in historesin. Also, methods for ATP-ase in samples for TEM.
Dear Doug, I suggest you look up articles by R.P. Apkarian. He has a Cr coating method which produces extremely fine grains. He told me of a detailed paper submitted to Scan. Microsc. Intl. which should have come out in 1994. I have a few of his papers, but not that one. Good luck. Yours, Bill Tivol
First Intarnational Conference on Electron Microscopy and Advances in Research in Differcent Fields of Sciance September 1995 Ismailia - Etap
Sponsored by Electron Microscopy Center Suez Canal University Ismailia - Egypt
Special Topics of the conference: 1- Role of EM in diagnostic virology. 2- Role of EM in diagnosis of tumors cylulogy and urinary atones. 3- Role of EM in ultrastructure pathology of the lung (non neoplastic conditions). 4- X-ray microanalysis: Applications particularly metalllurgical, mineralogical, and biological. 5- Scanning EM of plants, animal, ineccts, and mineral material. 6- Study of biological macromolecules from their characteristic electron diffraction patterns. 7- Skin pathology by EM. 8- Morphological ldentification of antigens by EM. 9- Different low temperature methods for biological EM. 10- Safety measures and maintenance needed for EM.
There will be an equipment exblbition in conjunction with this meeting. Registration for foreigners will be US $ 150 inclusive of full board during the time of the meeting. For further information, contact the organizar: Prof, Dr. Khalifa Ibrahim Khalifa Electron Microscvpe Center Suez Canal University Ismailia - Egypt Fax: (20) 64- 329478 ( phone and Fax number) Fax: (20) 64- 333318 ( Phone and Fax number)
Microwave irradiation has been shown to enhance the penetration of aldehyde into insect and plant tissues and to improve fixation of these specimens for LM and EM. Five references follow:
1. Lindley VA: A new procedure for handling impervious biological specimens. Microsc Res Tech 21:355, 1992
2. Smid HM, Schooneveld H, Meerloo T: Microwave fixation of water-cooled insect tissues for immunohistochemistry. Histochem J 22:313, 1990
3. Heumann HG: Microwave-stimulated glutaraldehyde and osmium tetroxide fixation of plant tissue: ultrastructural preservation in seconds. Histochem 97:341, 1992
4. Login GR, Dvorak AM: Methods of microwave fixation for microscopy. A review of research and clinical applications: 1970-1992. Prog Histochem Cytochem 27/4:1, 1994
5. Login GR, Dvorak AM: The Microwave Toolbook. A Practical Guide for Microscopists. Boston, Beth Israel Hospital Press, 1994, 184
In message {7247D4029F-at-marc.cri.nz} "IAN HALLETT" writes: } A message forwarded from my colleague Paul Sutherland } } Has anyone got a good fixation method for insect gut tissue which has } a thick peritrophic membrane. At present I am using 2% } paraformaldehyde, 2.5% glutaraldehyde in 0.1M phosphate buffer pH } 7.2 and a post fix in 1% osmium tetroxide. With this fix the } microvilli are not well preserved. } } } Ian Hallett
GRL glogin-at- bih.harvard.edu Beth Israel Hospital - Department of Pathology Telephone: 617-667-2034 Fax: 617-667-8676
Dear Listserver Audience: The tektronix image recording crt (Current # CRTC818P4S) on our Hitachi S-500 scanning EM is no longer functional. We have temporarily replaced it with a high resolution monitor, however this is not satisfactory due to the smaller image. Effort at expanding the image have resulted in some edge distortion and a loss of resolution. Does anyone out there have a replacement tube that they wish to sell? If so, please contact me at:
Dear Microscopy Listserver Audience: The tektronix image recording crt (#CRTC 818 P4S) on our Hitachi S-500 scanning EM is no longer functional. We have temporarily replaced it with a high res. monitor, however this is not satisfactory due to problems associated with the small image produced by the shorter yoke on this crt. Attempts at expanding the image electronically have resulted in some edge distortion and an unaccept- able level of resolution loss. Does anyone out there have a replacement crt of this type? If so, please E-mail me at: Bray-at-hg.uleth.ca
We have an 18 years old EDAX EDS system. The MCA and computer is slow and unreliable. Does anybody has information on upgrading old EDS systems by replacing the multichannel analyser and computer with an IBM PC or MAC? We would like to keep it as cheap as possible.
Thanks!
Kris
Kristof Kovacs Central Laboratory University of Veszprem Veszprem, P.O.Box 158 H-8201 Hungary Phone: +36-(88)-421684 Fax: +36-(88)-426016 e-mail: kris-at-miat0.vein.hu
Yes, you can upgrade old EDS systems. There is a program called WinEDS. Coupled with a plug in board for your PC, your detector and pulse processor you get a Windows based quantitative system.
Sorry I do not have the address of the person who wrote/design the system, however I will try and track him down (he is in Australia).
This may take a few weeks.
Keith Moulding ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ HKUST MCPC, Clear Water Bay, Kowloon Hong Kong.
I would suggest a call to 4-pi analysis. Their product line emphasizes upgrading existing EDS systems and works with NIH Image and DTSA (and others)
4-pi analysis 3500 Westgate Dr. Suite 403 Durham, NC 27707-2534 USA
phone: Mike Sczyz (919) 489-1757
fax: (919) 489-1487
electronic mail: go4pi-at-applelink.apple.com
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
The WinEDS system described by Keith Moulding is designed and marketed by Paul Thomson: Thomson Scientific Instruments 216 Drummond Street, Carlton, 3053, Victoria, Australia phone (03) 663 2738. fax (03) 663 3680
I had the opportunity to demo this system at the New Orleans MAS/MSA. It is an impressive package. This is not an endorsement, just a personal, scientific opinion.
This is a test. I tried to resend a message to this address and it was retu returned by the Rockefeller mail director. I couldn't resend it. Let's see if this gets out.
Seems to be a great disparity between the other packages I've looked at :
DTSA $800 and quirky Flame $7000 and Fast
Anybody know of any pakages somewhere in the middle? Center for Materials Research and Analysis Central Facility for Electron Microscopy University of Nebraska at Lincoln
I am trying to do fluorescent in situ hybridization with bacteria directly on Nuclepore polycarbonate membrane filters, and need to omit the alcohol series that is usually included in these protocols. Does anyone have any F.I.S.H. protocol that does not include the alcohol series? It doesn't need to be a membrane filter method. I am, however, interested in any F.I.S.H. membrane filter protocols. What types of membranes work best?
Thanks, Barry Pyle, Montana State University - Bozeman.
I don't know how thick you mean for the peritrophic membrane, but I've had good results by injecting fixative into the gut lumen and letting it work awhile before dissecting the gut out in fixative solution. This works well for lep guts. The fixative etc is described in Ryerse et al, Tissue and Cell, 24:751-771 1992. See discussion of preventing midgut cell surface blebbing on p 768 by injecting fix directly into the gut lumen. Good luck. Jan Ryerse, Pathology, St. Louis University Med School, St. Louis, MO
Message-Id: {MAILQUEUE-101.950327093814.384-at-vanlab.paprican.ca} To: microscopy-at-aaem.amc.anl.gov
Hi, For an economic EDX system set up to utilize your existing detector, I suggest you contact the following:
Dapple Systems, Sunnyvale CA , (408) 733-3283 They supply a PC or Mac based system which I believe is passive only( ie. does not take control of the beam)
4pi Analysis, Inc. Durham NC , (919) 489- 1757 They supply a Mac or Power Mac based system (with beam control?)
I have only read their brochures not used the systems, but I think they are worth a look.
Good luck, Laurie
Kristsf Kovacs wrote: } } We have an 18 years old EDAX EDS system. The MCA and computer is slow and } unreliable. Does anybody has information on upgrading old EDS systems by } replacing the multichannel analyser and computer with an IBM PC or MAC? We } would like to keep it as cheap as possible. } } Thanks! } } Kris } } Kristof Kovacs } Central Laboratory } University of Veszprem } Veszprem, P.O.Box 158 } H-8201 Hungary } Phone: +36-(88)-421684 } Fax: +36-(88)-426016 } e-mail: kris-at-miat0.vein.hu } }
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
A fully-funded position is available in my laboratory for a postdoctoral fellow or an advanced EM technician who can work independently. I am looking for someone with experience in thin sectioning of mammalian tissue culture cells and tissues. Specifically, this project involves immunoEM analysis by post-embedding procedures (e.g. Lowicryl or L.R. White) and/or ultrathin frozen sectioning. I am an Associate Professor at The Rockefeller University in New York. For those of you who may not know, Rockefeller is located in one of the best and safest areas of New York City. With neighboring Cornell Medical School and Sloan-Kettering Cancer Center, this is one of the most scientifically exciting communities to work in. Subsidized housing is available across the street, and child care is available on campus. My laboratory is studying endothelial adhesion molecules and their role in inflammation. We are taking a multidisciplinary approach using techniques of cell biology, immunology, molecular biology, and biochemistry. We have identified and cloned two unique adhesion molecules concentrated in the inter- cellular borders between endothelial cells. These are PECAM-1 (CD31) and the endothelial-specific cadherin, cad5 (VE-cadherin). Our lab has shown that PECAM is required for the migration of leukocytes through the endothelial junctions during inflammation. The project involves determining the ultrastructural distribution of these proteins along the endothelial junctions in vitro and in vivo. (We have also cloned the corresponding mouse PECAM and cad5.) We believe that part of the function of these molecules is determined by their distribution along the membrane. This would be an excellent oppportunity for someone trained in electron microscopy to further his/her skills as well as to learn new techniques of molecular biology and immunology. Our group interacts well and often, so that everyone can benefit from the others' knowledge and skills.
Interested parties should send a letter along with a C.V. to the address below. For specific questions or more details, please fax (212) 327-8875. I look forward to hearing from you.
Sincerely,
William A. Muller, MD, PhD Laboratory of Cellular Physiology and Immunology Box 176 The Rockefeller University 11230 York Avenue New York, NY 10021-6399
P.S. Thanks to Chris Jeffries and Jeanne Barker for helping me connect to this listserver.
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I have problems in embedding two types of material, one is developing oil seed rape pods in order to look at changes leading to pod shatter. Even after several days in 100% Spurr resin, the centre of the section, which often comtains the developing seed, remains soft and falls out when sectioning. Similarly, in a study on the infection of rubber leaves by colletotricum spp., when sectioning infected material the sections split along the leaf surface. Any ideas on overcoming these problems? I routinely use Spurr hard resin and go through propylene oxide after ethanol dehydration. I have also tried holding the specimens at 45C in the moulds for 24h before raising the temperature to 65C for full polymerisation. I have tried LRWhite with no improvement
Richard Pring mentioned: } I have problems in embedding two types of material, one is developing oil } seed rape pods in order to look at changes leading to pod shatter. Even } after several days in 100% Spurr resin, the centre of the section, which } often comtains the developing seed, remains soft and falls out when } sectioning. Similarly, in a study on the infection of rubber leaves by } colletotricum spp., when sectioning infected material the sections split } along the leaf surface. Any ideas on overcoming these problems? I routinely } use Spurr hard resin and go through propylene oxide after ethanol } dehydration. I have also tried holding the specimens at 45C in the moulds } for 24h before raising the temperature to 65C for full polymerisation. I } have tried LRWhite with no improvement
Very often the failure to infiltrate material adequately with resin is due to inadequate fixation and not to inadequate resin infiltration. Especially when processing oily samples it is necessary to fix {very} well to destroy all membrane semi-permeability and to stabilise the lipids. Going to extremes during the fixation may lead to substandard structural preservation, but may be the only way to adequately infiltrate the sample. One such overkill schedule is: Fix material 24h at 40deg C in 2.5% glutaraldehyde, then postfix for another 24h in several changes of osmium, also at 40deg C (take due care during this step). Dehydrate in acetone, use a few prop. oxide rinses and embed in your standard resin. Other methods (adding Malachite Green to aldehydes in DMSO) are also known and may very well work. All these methods have in common that they are not optimal as far as structural preservation is concerned, but sometimes they may be the only ones that will result in thin sections. Richard's second problem, that of the adhesion of the resin to the leaf surface is a result of the wax layer on the leaf surface forming a parting layer between the leaf and the resin. One solution is to use an epoxy formulation that is not as easily parted from cuticular waxes - Quetol may be a possibility.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
I have a an old Kevex EDS system hooked up to an "old" Cameca MBX microprobe....both about 15 years old. I've recently replaced the multi- channel analyzer with a 4pi processing system and a Mac computer. The 4 pi system costs around $6,000 and is easy to incorporate into many different EDS systems.
The 4 pi is relatively cheap, it's efficient and easy to operate and is capable of interfacing with many different EDS systems. Not only can it generate very nice EDS spectra through DTSA, it does an excellent job imaging elementally specific X-ray transitions. The quality of these EDS maps is close to that of the digital WDS maps that we generate on our SX-50 microprobe. It's really quite impressive.
If you are interested, I would be happy to FAX you some information on the 4 pi system as well as some EDS images we have generated using that system. Just let me know....
or here's 4 pi info:
4 Pi Analysis, Inc. 3500 Westgate Dr., Suite 403 Durham, NC 27707-2534 Tel: 919-489-1757 FAX: 919-489-1487
Has anyone had experience with long term storage of images from a thermal printer (Codonics, Seikosha, etc.)? I have noticed that the images start to fade away after exposure (months) to fluorescent lights. I have been told that this will happen even when they are protected from exposure to light. Any suggestions for storage or other experience would be appreciated.
John Giles Honeywell Space Systems (813) 539-2270 (813) 539-3630 Fax e-mail: jegiles-at-space.honeywell.com
We've had a Seikosha thermal printer for 4 years and still have some of the first images printed with it saved. The images have faded slightly from gray to brown, but you can only tell if you hold next to a new print. The best storage for them is in a binder or folder that is stored in a closet or file cabinet. As long as they aren't exposed to drastic variations in heat they should store for long periods of time with minimal fading.
------------------------------------------------------------------ Heinz Hemken Departamento de Biologia Celular, CINVESTAV-IPN http://cell.cinvestav.mx/bchh.html
Energy Dispersive Spectroscopy, which is probably more correctly called X-ray Energy Dispersive Spectroscopy (XEDS) but various other rearrangements of the letters exist in the literature (EDXS,...)
It is an analytical methodology in which a solid state semi-conducting detector (either Si or Ge based) is used to measure the energy distribution of X-rays emitted from a specimen. The X-ray's may be excited by some sort of probe (usually a focussed beam of electrons, ions, or photons). The "Spectrum" is then displayed on a graphical output device (read computer screen) and the relative intensity of the measured characteristic X-ray peaks, analyzed to determine the relative elemental composition of the material.
Any good text book on Scanning Electron Microscopy or Electron Probe Microanalysis (published within the last 10 years or so will have a chapter or two on the subject).
An article about 4pi system for EDS was published in SCANNING Vol. 14, 233-240 (1992). The title is:
Mac X-ray: The NIST 'DTSA' program and the 4pi analysis board.
one of the authors is: David C. Joy EM lab U. of Tennessee Knoxville, TN 37996-0810
Our lab also installed both the 4pi spectral engine boards and the 4pi scanning interface box. We have been enjoying NIH Image and several other programs in playing the SEI, BEI, and X-ray mapping.
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
As promised, following is the summary of responses received from my question, "How many of you have your curing ovens vented to a hood?".
From a total of 11 responses, one containing knowledge of two others, there were only two who were not curing their resins in, or venting their ovens to a hood. Both are making provisions to do so. Nine actually place their ovens in the hood while two are vented to the hood. There was only one reference sited...that being an article in the Proceedings of the RMS Vol. 16, pp. 265-270, 1981. Most of us seem to subscribe to the "just to be sure" school of reason. I attempted to thank each of you who responded individually, but I think that I may have lost one of you. So let me say now....thank you very much.
Sandra Zane Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
The original paper on PLP was McLean and Nakane 1974, J Histochem Cytochem 22:1077-1083. Also see Pino 1984, Stain Technol. 59:307.
Some of the most impressive uses of PLP came out of Marilyn Farquhar's lab. If I remember correctly the ICC that Farquhar/Brown did utilized PLP, and would be a good source of method. For example, see: Cell 36:295(1984). PNAS 81:5135 (1984). JCB 106:1863 (1988). Meth Cell Biol 31:553 (1989).
Good luck.
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School Ann Arbor, MI 48109-0616 {akc-at-umich.edu}
---------------------------------
On Mon, 27 Mar 1995, Michel Deschuyteneer wrote:
} I would greatly appreciate the reference and/or the recipe for the } Paraformaldehyde - Lysine - Periodate (PLP) fixative. } Thank you very much in advance. } } Regards, } Michel. } Michel Deschuyteneer deschuyt-at-sbbio.be } Scientist - Electron Microscopy Laboratory } SmithKline Beecham Biologicals } Rue de l'Institut, 89 - B1330 Rixensart, BELGIUM } Tel: +32-2-656 9290 Fax: +32-2-6568113 } } } }
I have a an old Kevex EDS system hooked up to an "old" Cameca MBX microprobe....both about 15 years old. I've recently replaced the multi- channel analyzer with a 4pi processing system and a Mac computer. The 4 pi system costs around $6,000 and is easy to incorporate into many different EDS systems.
The 4 pi is relatively cheap, it's efficient and easy to operate and is capable of interfacing with many different EDS systems. Not only can it generate very nice EDS spectra through DTSA, it does an excellent job imaging elementally specific X-ray transitions. The quality of these EDS maps is close to that of the digital WDS maps that we generate on our SX-50 microprobe. It's really quite impressive.
If you are interested, I would be happy to FAX you some information on the 4 pi system as well as some EDS images we have generated using that system. Just let me know....
or here's 4 pi info:
4 Pi Analysis, Inc. 3500 Westgate Dr., Suite 403 Durham, NC 27707-2534 Tel: 919-489-1757 FAX: 919-489-1487
I have a an old Kevex EDS system hooked up to an "old" Cameca MBX microprobe....both about 15 years old. I've recently replaced the multi- channel analyzer with a 4pi processing system and a Mac computer. The 4 pi system costs around $6,000 and is easy to incorporate into many different EDS systems.
The 4 pi is relatively cheap, it's efficient and easy to operate and is capable of interfacing with many different EDS systems. Not only can it generate very nice EDS spectra through DTSA, it does an excellent job imaging elementally specific X-ray transitions. The quality of these EDS maps is close to that of the digital WDS maps that we generate on our SX-50 microprobe. It's really quite impressive.
If you are interested, I would be happy to FAX you some information on the 4 pi system as well as some EDS images we have generated using that system. Just let me know....
or here's 4 pi info:
4 Pi Analysis, Inc. 3500 Westgate Dr., Suite 403 Durham, NC 27707-2534 Tel: 919-489-1757 FAX: 919-489-1487
Return-Path: u096585-at-mdanl3.md.dow.com Received: by mdanl3.md.dow.com (UCX V2.0-15) Wed, 29 Mar 1995 08:22:41 -0500 Received: from aaem.amc.anl.gov by inet1.ma.dow.com with SMTP id AA10523 (InterLock SMTP Gateway 1.1); Wed, 29 Mar 1995 08:22:24 -0500
Thanks to all who have explained to me what EDS is. I got several enlightening and courteous replies. This is a great list and a great community!
Keep up the rich info stream!
------------------------------------------------------------------ Heinz Hemken Departamento de Biologia Celular, CINVESTAV-IPN http://cell.cinvestav.mx/bchh.html
He is using SEM for industrial applications (various analyses of Clay, paints, paper, etc). They are looking for a good short course on SEM in materials science. The dates of both the Lehigh and McCrone courses will not fit into their schedule. I know of a course at Univ Michigan which I told them about. Are there any others?- Thanks for any help you can render.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
I am trying to do in situ hybridization and immunohistochemistry on glycol methacrylate-embedded wheat pollen. The reason I am using that resin, despite its uglyness to work with, is that the infiltration of pollen is very good and the overall morphology seems to be well preserved. But to gain a better access to the target in the tissue I would like to remove the resin after sectionning. The resin was polymerized with benzoyl peroxide at 55C. Is there a way to remove it without affecting antigenicity of the tissue???
If I cannot remove the glycolmethacrylate, I am planning to use a mixture of butyl and methyl methacrylate with a polymerization under UV. According to litterature, I can remove it with acetone. Some people use benzoin, others use benzoin ethyl ether; what is the difference between the two? Can I use bezoyl peroxide instead? I know that UV polymerization can lead to inconsistancy in the polymerization, is the use of a 4W placed on top of the specimen at a distance of about 10 cm could be good?
Since I do not subscribe to this group could you send me directly any suggestions you may have.
Greetings: I'm interested in using fluorescent probes to clarify the relationship between certain cellular structures, specifically the ER and what I believe to be vacuoles. Some of my colleages have suggested that the vesicles I believe to be vacuolar in origin may be dilated ER. In looking for likely probes I've found refences in the Molecular Probes handbook to DiIC (a long-chain carbocyanine) and Rhodamine B as ER-specific dyes. However, the cited references are based on experiments on animals. Ideally, I would like to use two probes that will discriminate between ER and vacuoles. Does anyone have experience with these dyes or could someone point me towards some references? I'm working with leaf tissue, specifically cells with the minor veins. Many thanks in advance.
Dwight Beebe IRBV, Dept. de sciences biologiques beebed-at-ere.umontreal.ca Universite de Montreal Voice:514-872-4563 4101, rue Sherbrooke est FAX:514-872-9406 Montreal, PQ H1X 2B2 Canada
Dear friends, Thanks for your responses. As you all may remember I was having some crossing over problem between FITC and Rhodamine-that's what I thought... My problem seems to be autoflorescense, yeah that simple and basic, I just overlooked it. I just looked at the neurons with just plain solution, they flouresce beautifully under the rhodamine filter, and slightly under the FITC filter and no fluorescence under the Cy5 filter. Also, I had a control (no antibodies present) which I fixed with 4% paraformaldehyde and used 3% normal goat serum as blocking reagent. This control flouresce better under the FITC filter than under the Rhodamine filter, the opposite of the control without fixative. It does not flouresce under the Cy5 filter. Also, I did a control with CY5 and Cy3. Again the cells flouresces under the FITC and rhodamine Filters but not under the CY5 filter. So my solution seems to stay away from FITC, and rhodamine. Any suggestion on how I can better solve my autoflourescence problem will be appreciated. Thanks again for your responses. Your friend, Ciprian
__________________________________________________________ Ciprian Almonte Medical College of Pennsylvania Dept. of Anatomy and Neurobiology E-mail: almonte-at-medcolpa.edu 3200 Henry Ave. Voice: (215) 842-4081 Philadelphia, PA 19129 Fax: (215) 843-9082 __________________________________________________________
There is a very good SEM short course given by Botany department of university of washington in Seattle. This course covers various subject and the scheduel is flexable. The instructor of the course is Barbra Reine.
Tel # for Botany Department: (206) 543-1942
On Wed, 29 Mar 1995, Jay Jerome wrote:
} This request is for a colleague: } } He is using SEM for industrial applications (various analyses of Clay, } paints, paper, etc). They are looking for a good short course on SEM in } materials science. The dates of both the Lehigh and McCrone courses will } not fit into their schedule. I know of a course at Univ Michigan which I } told them about. Are there any others?- } Thanks for any help you can render. } } } Jay Jerome } ************************************************************** } * aka: W. Gray Jerome * } * Dept. of Pathology * } * Bowman Gray School of Medicine of Wake Forest University * } * Medical Center Blvd * } * Winston-Salem, NC 27157-1092 * } * 910-716-4972 * } * jjerome-at-isnet.is.wfu.edu * } ************************************************************** } }
If someone knows, please, write me, which type of NCAM antibody can work for post- embedding method? (Araldite 6005 or Epon resin is used and pre embedding I have made an enzyme histochemical reaction. Fixation : paraformaldehyde- glutaraldehyde in cacodylate buffer) Thanks! Agnes Kittel
Both my lab and another lab here in Hawaii have Denton DV-502 vacuum evaporators. My evaporator is perfectly fine for my needs; I can carbon coat Formvar grids or prepare pure carbon substrates for TEM. The other lab, however, wants to carbon coat for SEM, with and without EDS. Their problem is that they cannot get the carbon to evaorate long enough to get a thick enough coating, and neither can I on my instrument. We both have the same spring-steel (?) driven carbon rod holders (which differ from the gravity-fed system in the very old Denton in still another Hawaii lab and also from the coiled spring feed on the non-adjustable holders). The problem seems to be that there is enough resistance somewhere that everything heats up like crazy, the moveable parts expand enough to get stuck, and the whole feed process stops until the unit cools down or blows a fuse, whichever comes first. This has happened with old holders, brand-new ones, and everything else I have been able to devise. Has anyone else had and oversome this problem? Any hints and tips will be appreciated!
Mahalo and aloha, Tina Weatherby Carvalho Biological EM Facility University of Hawaii
We need information on where to get a light scrambler for homogeneous illumination -to couple an HBO light source to a Zeiss microscope for fluorescence work. Sources, FAX numbers, price ranges -any information would be highly appreciated. dd
Daniel Dagan, Dept. Biophysics and Physiology, Faculty of Medicine, Technion, Israel Institute of Technology, POBox 9697 Haifa 31096 ISRAEL
From your description, it sounds like you need quenching rather than more blocking. Blocking uses miscellaneous protein to block non-specific protein-binding sites in the tissue, but doesn't actually deal with autofluorescence. Quenching uses reducing agents such as ammonium chloride or sodium borohydride to reduce double bonds in the tissue, which are usually the main source of autofluorescence. Try the following: make up a fresh 1% solution of sodium borohydride in distilled water (careful, borohydride is toxic), and leave a few drops of it on your sections for about 10 minutes early in your immunocytochemical run (before the primary antibody step). This should reduce the inherent autofluorescence in your tissue.
A. Kent Christensen, Department of Anatomy and Cell Biology, University of Michigan Medical School, {akc-at-umich.edu}
------------------------------------
On Wed, 29 Mar 1995 almonte-at-medcolpa.edu wrote:
} Dear friends, } Thanks for your responses. As you all may remember I was having } some crossing over problem between FITC and Rhodamine-that's what I } thought... My problem seems to be autoflorescense, yeah that simple and } basic, I just overlooked it. } I just looked at the neurons with just plain solution, they } flouresce beautifully under the rhodamine filter, and slightly under the } FITC filter and no fluorescence under the Cy5 filter. Also, I had a control } (no antibodies present) which I fixed with 4% paraformaldehyde and used 3% } normal goat serum as blocking reagent. This control flouresce better under } the FITC filter than under the Rhodamine filter, the opposite of the } control without fixative. It does not flouresce under the Cy5 filter. } Also, I did a control with CY5 and Cy3. Again the cells flouresces } under the FITC and rhodamine Filters but not under the CY5 filter. } So my solution seems to stay away from FITC, and rhodamine. Any } suggestion on how I can better solve my autoflourescence problem will be } appreciated. Thanks again for your responses. } Your friend, } Ciprian } } __________________________________________________________ } Ciprian Almonte } Medical College of Pennsylvania } Dept. of Anatomy and Neurobiology E-mail: almonte-at-medcolpa.edu } 3200 Henry Ave. Voice: (215) 842-4081 } Philadelphia, PA 19129 Fax: (215) 843-9082 } __________________________________________________________
A message about an article regarding the 4pi system sent out couple of days ago was bounced back. The paper was published in SCANNING Vol. 14, 233-240 (1992). The title is:
Mac X-ray: The NIST 'DTSA' program and the 4pi analysis board.
One of the authors is: David C. Joy EM lab U. of Tennessee Knoxville, TN 37996-0810
Our lab has installed both the 4pi spectral engine boards and the 4pi scanning interface unit. We have also been enjoying NIH Image and several other programs in playing the SEI, BEI, and X-ray mapping.
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
} Does anyone know the mechanism of geode crystal formation in the midwest? And } why do some geodes contain oil?
According our resident geologist, geodes form when percolating waters precipitate into hollow cavities. Whatever happens to be carried with the water is deposited in layers into the cavity (this includes oil).
1. Has anyone made comparison about the performance, life time, and potential contamination between new filament and rebuilt one. The prices are four times difference. 2. Which company supplies better batch of filaments. This information may mail to me directly if you feel it may offend some one. 3. Last year, we had a batch of new filaments that tend to form thin film of metal sticking on the tip of the filament, though the heating temperature was low (with slightly large distance between filament and the cap, and the heating current is limited by a stopper). Is this a problem due to impurity of filament? Any idea?
Thanks for any input.
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
Tina: In our lab, the DV-502 is used for C coating SEM and Microprobe samples for EDS and WDS works.
1. The length and size of the reduced C rod are important factors for the coating thickness. The coarser C rod does not only supply more C but also support longer reduced C rod. However, coarser C rod needs higher current to reach the evaporation temperature, hence it might hit some limits of the machine, like burn the fuses. This has to be balanced. 2. You also need to properly control the heating current during evaporating. Specially, you need quickly bring the current back to some position where evaporation can last longer at the very beginning when the evaporating starts.
Xiaogang
**************************************** * Xiaogang Xie * * Department of Geology and Geophysics * * Louisiana State University * * Baton Rouge, LA 70803 * * Office (504)388-2240 * * Fax (504)388-2302 * ****************************************
Message-Id: {m0ruVG4-0007AdC-at-stjohns.ohsu.edu} Message-Version: 2 } To: microscopy-at-aaem.amc.anl.gov
I have colleague that needs help setting up an automatic computer based method to count gold particles on TEM micrographs. He has NIH image, a good Macintosh and can import digital images but is having difficulties getting accurate counts of the gold particles. If anyone has experience with this please let me know.
Subject: Time: 4:13 PM OFFICE MEMO RE} C evap for EDS Date: 3/30/95
Tina- check your ceramic insulators through which the rod holder slides. If the ceramic gets enough carbon or metal on it, it creates a conductive pathway which corrupts your normal electrical flow and sends current through the metal parts of the electrode instead of the carbon rod. Could explain the hot metal and blown fuses from over-amping the system. This can even melt some of the smaller metal parts. Cleaning the ceramic is tough because of the porous surface and chances are the ceramic will crumble from the excessive heating when you dissassble the holder. Suggest you order some new ones from Denton and keep some spares. Have you tried the carbon yarn yet? Works well, never sparks or arcs, heavier thicknesses can be produced by reducing the source to target distance. Last time I checked, Denton had the best price. -Doug
We have done a comparison study of rods from different sources, and found some to be unworkable for SEM/EDS coating in our 502. Rather than belittle the ones that don't work, I can recomment those from Ladd. Hope this helps.
Subject: Time: 9:28 AM OFFICE MEMO Denton evap Date: 3/30/95
Tina- We have a 502A with turbo. One thing to check immediatly is the ceramic insulators which the electrode holder slides through. In the past, repeated applications of heavy coatings will eventually create a conductive surface coating on these parts and current through the carbon rod will be reduced as it flows along this alternate pathway. In extreme cases, carbon will not evaporate and metal parts of the electrode will actually begin to melt. Good thing your fuses are blowing rather than your top. The insulators are tough to clean because of the porous surface and you may find that they crumble to dust when you remove them due to the heating they have been subjected to. It's good to have a few extras around as this is a chronic problem. Have you tried the carbon yarn yet in the Denton? It is impossible to blowout/arc as with the rods and is much more controllable, just evaporate until it burns through. Heavier coatings can be acquired by reducing the distance from the source. Look out for metal contamination of the yarn if you evaporate shadowing metals; it shows up as tiny electron-dense spots on you substrates. Use the shutter to protect the unused yarn on the bobbin. You can get the yarn directly from Denton for the best price; e-mail me if you need info. -Doug
Message-Id: {MAILQUEUE-101.950330104455.320-at-vanlab.paprican.ca} To: microscopy-at-aaem.amc.anl.gov
Hello, We are intending to purchase a carbon evaporation unit for coating SEM/EDX samples . We have received quotes for 6 systems but have narrowed the choises to the following on the basis of cost: Denton 502A Ladd Edwards 306 The price range for these units is $24,000-$35,000 in Canadian dollars. We are very tight for money (of course) but do not want to buy a unit which will give us continuous headaches with overheating, jamming of the carbon rod feed mechanism or having to evacuate several times in order to recoat because enough carbon could not be applied the first time. If you have experience with any of these units, either positive or negative, I would appreciate hearing your comments. If you have any experience with using 1/4" rods vs. 1/8" rods or "reduced section" versus conically tipped rods, please pass it along.
I was all set to make a purchase before I read the recent comments on the listserver. Thanks very much for any assistance. Laurie
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
Message-Id: {MAILQUEUE-101.950330143551.320-at-vanlab.paprican.ca} To: microscopy-at-aaem.amc.anl.gov
Hi, I am trying to wrap up a summary of referances on EDX and require some information: 1) Has the book from MAS entitled "X-ray Spectrometry in Electron Beam Instruments" by Dave Williams and Dale Newbury actually been published yet and if so, is there a phone number for orders?
2) Is anyone familiar with an EDX upgrade system from "Aptec"? I believe it is located somewhere in Ontario, Canada but I am not sure if the name I have is a company or product name. Contact # ?
3) When and where is the next International Congress on Electron Microscopy (ICEM) and what is a contact number for information?
4) When and where is the next International Conference on X-ray Optics and Microanalysis (ICXOM) and what is a contact # for info?
I will be making a summary of the information I have gathered on EDX available shortly. Thanks for any assistance. Laurie
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
Reply to: RE} TEM: Gold particle counting Hi Bob, Your colleague may be suffering from low contrast images. Try acquiring or scanning in the images with greater contrast range. The contrast of the images should be such that the black gold particles should be black (an extreme gray level) and everything else in the image should be a different gray value (mid-gray to white). For the images that are already digitized try doing a "Sharpen" or an "Unsharp Masking" operation on the image.
Sincerely,
George McNamara Universal Imaging Corporation --------------------------------------
I have colleague that needs help setting up an automatic computer based method to count gold particles on TEM micrographs. He has NIH image, a good Macintosh and can import digital images but is having difficulties getting accurate counts of the gold particles. If anyone has experience with this please let me know.
Message-Id: {9503311445.AA01865-at-dnagel.bio.fc.ul.pt} X-Sender: bjfeijo-at-skull.cc.fc.ul.pt X-Mailer: Windows Eudora Version 1.4.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I there: One of our users has plans to look at entire styles and to do some immuno labeling of the stylar channel proteins. He wants to get rid of embedding and/or freeze-sectioning so he figured confocal would be the answer. Among others, I antecipate some problem with self fluorescence coming either from chlorophyls and/or lignins. One way around these would be to use the red line of the Kr-Ar (we have a MRC-600). So, does anyone has experience on working with secondary antibodies (in this case against chicken) labeled with red excitable fluorophores in plant tissues? What may the major drawbacks in this kind of preparation? Any help would be highly appreciated.
___________________________________________________________ Jose A. Feijo Dept. Biologia Vegetal, Fac. Ciencias Lisboa Ed. C2, Campo Grande, P-1700 LISBOA, PORTUGAL
The Carnegie Mellon Research Institute (a division of Carnegie Mellon University) is trying to find a new home for a 13 year old ISI SS-40 SEM. If anyone out there is interested please send mail to me at bz0c+-at-andrew.cmu.edu
} From the Characterization Facility at the University of Minnesota
MASTER CLASS
Cryo-TEM of Colloidal Materials
=A5 Specimen Preparation
=A5 Cryo-Transfer to the Electron Microscope
=A5 Low-Electron Dose Image and Diffraction Pattern Recording on Film
=A5 Image-Processing and Analysis of Digital Cryo-TEM Images
May 18-20, 1995
Intended Audience
This Master Class is intended for any materials scientists or process eng= ineers =
working with biological or colloidal materials. This class will explore t= he use =
of the technique of cryo-transmission electron microscopy to probe the =
microstructure of these samples in the vitrified, hydrated state at resol= utions =
around 1 nm. Previous TEM experience, although welcome, is not required.=
Description
The lectures will introduce the basics of cryo-transmission electron micr= oscopy, including sample preparation and microscopy techniques. Applications to s= pecific biological and colloidal materials=D5 samples will be demonstrated, and =
complementary characterization techniques will be described.
The details of sample preparation, focussing on vitrification parameters = and =
transfer to the microscope, will be described and demonstrated. Imaging =
techniques and interpretation, artifact identification, and quantitative =
analysis will also be covered.
Supervised hands-on laboratories demonstrating the techniques that were =
introduced in the lectures will be interspersed throughout the class. Po= ssible =
topics include: low dose imaging of soft latex particles; low dose imagi= ng of =
phospholipid vesicles; image processing of cryo-TEM images using NIH Imag= e and =
Digital Micrograph; CEVS sample preparation; and video-enhanced light mic= roscopy of colloidal materials.
Upon completion of the class, participants will be able to select an appr= opriate cryogen, prepare vitrified thin films of colloidal specimens, transfer th= ese =
specimens to the microscope, and record images free from artifacts caused= by =
electron beam damage.
Registration
For more information, reply to Beth Trend, trend-at-cems.umn.edu
Instructors
Frank Booy is a Senior Staff Fellow, National Institute of Arthritis =
Musculoskeletal and Skin Diseases at the National Institutes of Health. = As a =
special expert in electron microscopy, he has worked in the field of =
cryo-electron microscopy of biological materials since 1978, at the CNRS = in =
Grenoble, France, the European Molecular Biological Laboratory in Heidelb= erg, =
Germany, and the Rijksuniversiteit Groningen in the Netherlands. Dr. Boo= y is =
particularly involved in the use of cryo-electron microscopy and 3-D imag= e =
reconstruction techniques to localize the proteins that make up the =
nucleocapsids of herpes simplex virus.
John Minter, a Research Associate in the Analytical Technology Division o= f the =
Eastman Kodak Company, is the technical group leader of the Quantitative = Colloid Microscopy Group. Minter=D5s group applies cryo-TEM and electron crystall= ography =
to the study of photographic colloids. During 1990, he was an Industrial = Fellow =
at CIE collaborating with Professors H. Ted Davis and L. E. Scriven to st= udy =
surfactant morphology and crystal precipitation by cryo-TEM. He is curren= tly an =
Adjunct Professor at CIE and Industrial Co-chair of the CIE Characterizat= ion =
Facility Advisory Committee.
David P. Siegel is a Senior Scientist in the Research & Development Dept.= , =
Procter & Gamble Co. His general interest areas are membrane biophysics = and the physical chemistry of biologically relevant lipids. Dr. Siegel is partic= ularly =
interested in the mechanisms of biomembrane fusion and of lamellar-to-inv= erted =
phase transitions in phospholipids. Since 1991, he has used cryo-TEM and= =
time-resolved cryo-TEM to image intermediates in these processes.
Schedule
Thursday May 18 =
8:00 a.m. Registration =
8:30 Lecture I: Introduction to Cryo-TEM =A5 Basics steps in cryo-TEM =A5 Biological and colloidal problems that can be solved using c= ryo-TEM =
=
10:15 Lecture II: Sample Preparation, Vitrification, Storage, and Trans= fer to =
the Microscope =
Lunch =
1:00 p.m. Lab Session - Shepherd Labs
Dinner
7:00-10:00 Lab Session continues as needed =
Friday, May 19
8:00 Coffee =
8:30 Lecture III: Imaging in Cryo-TEM =A5 Operation of Electron Microscope =A5 Recognizing the good samples to examine =
10:15 Lecture IV: Image interpretation, processing, and analysis =A5 Artifacts - a rogue's gallery =A5 Quantitative analysis
____________________________________________________________________= ___ Beth Trend trend-at-cems.umn.edu (faster) or btrend-at-maroon.tc.= umn.edu Coordinator, Characterization Facility University of Minnesota Center for Interfacial Engineering =
100 Union St SE Room 187 phone 612-624-1365 fax 612-626-7530=
The Carnegie Mellon Research Institute (a division of Carnegie Mellon University) is trying to find a new home for a 13 year old ISI SS-40 SEM. If anyone out there is interested please respond to me at bz0c+-at-andrew .cmu.edu
In our newsletter, Microscopy Today, we attempt to publish monthly an on-going summary of ALL microscopy events (conferences/meetings, schools, training sessions, etc.) in the U.S. - plus major international events. A no-charge listing includes: Dates Title Sponsor/Organizer Location Contact/Tel/Fax We will consider longer write-ups and ask for an article contribution to the newsletter, rather than money, for the effort. The newsletter is mailed only to those who have requested copies - at no charge currently to some 8,000 microscopists in the U.S., Canada and the U.K. Unfortunately we must charge a modest subscription for other international readers. We invite your no charge event listing - or request for a subscription. Regards, Don Grimes, Editor
We would be obliged if you informed us about the specifications (including ISBN codes) of those books/atlases/workbooks which were published for students in the field of ULTRASTRUCTURAL HUMAN PATHOLOGY in the last ten years. Could you recommend for us some? Thank you in advance,sincerely yours
First Intarnational Conference on Electron Microscopy and Advances in Research in Differcent Fields of Sciance September 1995 Ismailia - Etap
Sponsored by Electron Microscopy Center Suez Canal University Ismailia - Egypt
Special Topics of the conference: 1- Role of EM in diagnostic virology. 2- Role of EM in diagnosis of tumors cylulogy and urinary atones. 3- Role of EM in ultrastructure pathology of the lung (non neoplastic conditions). 4- X-ray microanalysis: Applications particularly metalllurgical, mineralogical, and biological. 5- Scanning EM of plants, animal, ineccts, and mineral material. 6- Study of biological macromolecules from their characteristic electron diffraction patterns. 7- Skin pathology by EM. 8- Morphological ldentification of antigens by EM. 9- Different low temperature methods for biological EM. 10- Safety measures and maintenance needed for EM.
There will be an equipment exblbition in conjunction with this meeting. Registration for foreigners will be US $ 150 inclusive of full board during the time of the meeting. For further information, contact the organizar: Prof, Dr. Khalifa Ibrahim Khalifa Electron Microscvpe Center Suez Canal University Ismailia - Egypt Fax: (20) 64- 329478 ( phone and Fax number) Fax: (20) 64- 333318 ( Phone and Fax number)
First Intarnational Conference on Electron Microscopy and Advances in Research in Differcent Fields of Sciance September 1995 Ismailia - Etap
Sponsored by Electron Microscopy Center Suez Canal University Ismailia - Egypt
Special Topics of the conference: 1- Role of EM in diagnostic virology. 2- Role of EM in diagnosis of tumors cylulogy and urinary atones. 3- Role of EM in ultrastructure pathology of the lung (non neoplastic conditions). 4- X-ray microanalysis: Applications particularly metalllurgical, mineralogical, and biological. 5- Scanning EM of plants, animal, ineccts, and mineral material. 6- Study of biological macromolecules from their characteristic electron diffraction patterns. 7- Skin pathology by EM. 8- Morphological ldentification of antigens by EM. 9- Different low temperature methods for biological EM. 10- Safety measures and maintenance needed for EM.
There will be an equipment exblbition in conjunction with this meeting. Registration for foreigners will be US $ 150 inclusive of full board during the time of the meeting. For further information, contact the organizar: Prof, Dr. Khalifa Ibrahim Khalifa Electron Microscvpe Center Suez Canal University Ismailia - Egypt Fax: (20) 64- 329478 ( phone and Fax number) Fax: (20) 64- 333318 ( Phone and Fax number)
Dear Szabolcs and Evelin, I have several recommendations for you (though I'm sorry I don't have the ISBN codes handy). I would recommend the Journal: Ultrastructural Pathology. I would also recommend becoming a member of the Ultrastrucural Pathology Society. This society has among its members some of the "giants" in this field. To join, or inquire for more information please contact:
Claire M. Payne, Ph.D. Secretary, Ultrastructural Pathology Society University of Arizona Dept. of Microbiology & Immunology 1501 N. Campbell Tucson, AZ 85724 USA
Books:
Diagnostic Ultrastructure of Non-Neoplastic Diseases (1992), Papadimitriiou, Henderson and Spagnolo, Churchill Livingstone, Pub.
Diagnostic Electron Microscopy: A Text/Atlas (1988), Dickerson, Igaku-Shion, Pub.
} We would be obliged if you informed us about the specifications } (including ISBN codes) of those books/atlases/workbooks which were } published for students in the field of ULTRASTRUCTURAL HUMAN PATHOLOGY in } the last ten years. } Could you recommend for us some? } Thank you in advance,sincerely yours } } Szabolcs Viragh and Evelin Orso } }
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (520)626-2824 dcromey-at-ccit.arizona.edu
I see several folks trying to subscribe to the Fatfree list using the wrong address. This is the subscription address: FATFREE-REQUEST-at-HUSTLE.RAHUL.NET
To join, send e-mail to the subscription address using one of the following subjects: ADD to join as a regular member ADD DIGEST to join as a digest member
Hope this helps.
At 12:01 PM 4/3/95, JONES-at-KRDC.INT.ALCAN.CA wrote: } Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
Dear Laurie, We have an old Ladd and a couple of home-made evaporators. The Ladd has been here longer than I (15 years), gets heavy use, and still works just fine (we just use it for coating grids, etc., so your mileage may vary). We use the 1/8" rods narrowed to ~1 mm (reduced section). I have coated objects from barely visible to very dark in one shot. BTW, a piece of paper folded so that one part shadows the other allows the color of the carbon to be fol- lowed relatively easily once one gets the hang of it. Any bell jar can, of course, be combined with the evaporating system (power supply, rod holder, etc.), so if you have the pumps available, you might save some $ by buying parts. Good luck. Yours, Bill Tivol
Dear Xiaogang, We have used both new and rebuilt W filaments, and have found the re- built ones to be at least as good as the new ones--in some ways better. The rebuilt ones we bought were from EBTEC (I am only a customer & have no finan- cial interest). The advantages of EBTEC's designs are 1) the connection of the hairpin to the posts is better than on some other designs, and 2) they have "regular" and "sharp" tips. The regular tips give a nice bright beam, and the sharp ones give a smaller source size. The intensity per unit area for the sharp tips is about the same as that for the regular tips, so the total inten- sity is less; however, the coherence is better, so for diffraction with radia- tion-sensitive specimens, the sharp tips are very good. We routinely preheat our filaments before mounting them in the Wehnelt cylinders. I don't know what the thin film you find sticking to your filaments is, but maybe preheating will help. Good luck. Yours, Bill Tivol
With 20 years of experience in servicing DV502's the most common problems I have encountered with carbon evaporation are:
1. Early units did not have a braided conduction wire between the moveable collet and the frame for sure current conduction. The fix for this is get the braid and drill/tap holes for fastening. If this is a problem, contact Denton to send it in for upgrade.
2. All versions work well with reduced section rods (approx .7mm diameter), pointed rods vary in conduction from start to finish and when the size nears the full 2mm the current can well go over 50 amps. I find reduced section rods will evaporate between 20-30 amps.
3. If the fuse is blown, often the fuse is replaced with a standard fuse. The correct fuse would be a ceramic type ABC fuse which is more tolerant and safer when it blows.
4. With reduced section rods if evaporation is continued past the reduced section you get the same overcurrent as with pointed rods.
5. Unfortunately there is a lot of variance in the way carbon is made into rods. You will see a variance of color, brittleness, ash formation (from some filler!). Only buy small quantities of any type until you find one that is good.
Obviously constant pressure on the rods during evaporation is essential. The leaf spring used for pressure can be easily abused by pulling it straight back to put in carbon rods. A better way of displacing it is to push it down for loading the carbon rod. If there is poor tension you really have to remove the spring and reform it to put more pressure on the rod.
7. If all of this doesn/t help, call the company and GET HELP, there is no reason to put up with poor performance, all parts and advice are available. The phone at Denton is 609-439-9100.
For any more detailed discussions you may contact me at Integrated Microsystems, Inc. 708-698-4210, fax 708-696-2541 or email.
I don't want to sound like I'm beating down the Egyptian meeting. But we've had multiple postings of that announcement for the last few weeks. I think we've had enough. Unfortunately, they are from different sources every time. So I cannot touch base with one person/organization and ask them to stop. So as a general announcement
Please cease posting this announcement until there is a significant change!
I remember seeing a request in the last few weeks about diamond knives. We are about to purchase one and are looking for suppliers and any thoughts on who makes the best diamond knives. I seem to remember the last time I bought one that the best were from Dupont or Diatome(?) but can't remember exactly which. Is that still true.? Any help greatly appreciated.
I've had a similar problem with this list that I haven't had with others. My postings get to the list, but somewhere out there multiple copies bounce off servers and come back over a period of days. Not being a UNIX guru, I have no idea why this is.
------------------------------------------------------------------ Heinz Hemken Departamento de Biologia Celular, CINVESTAV-IPN http://cell.cinvestav.mx/bchh.html
On Mon, 3 Apr 1995, Nestor J. Zaluzec-Argonne Nat. Lab. wrote:
} Colleagues... } } I don't want to sound like I'm beating down } the Egyptian meeting. But we've had multiple } postings of that announcement for the last few weeks. } I think we've had enough. Unfortunately, they } are from different sources every time. So I cannot } touch base with one person/organization and } ask them to stop. So as a general announcement } } Please cease posting this announcement until } there is a significant change! } } Thanks... } } Nestor } Your Friendly Neighborhood SysOp. } } } }
X-Mailer: InterCon TCP/Connect II 2.1.2 Mime-Version: 1.0 Message-Id: {9504032201.AA06079-at-usd14716.interramp.com}
I am working with sea urchin eggs and would like to have pictures of untreated eggs plus treated eggs ( with DTT and alpha-amylase) with SEM and TEM. Iam not sure what fixatives are appropriate for SEM and TEM . Please help me.
I did a considerable amount of TEM and SEM on urchin eggs a number of years ago. As I recall, the fixative giving the best results was something like 2% Glut. in seawater followed by osmium post-fixation. For references, see Byrd and Eppel, and Belisle and Byrd from the period of 1974 to 1979. Sorry I can't be more specific.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
Good Morning Microscopists, Yesterday morning after observing several folks trying to subscribe to this list using the wrong address and some folks attempting to do the same with another list, I thought I would be helpful. Well....I somehow got my wires crossed and it seems that a great many of you now know how to join that other list. Please accept my apologies and I promise that I shall never again attempt to be helpful on a Monday morning. (Hopefully, I provided a chuckle for some of you.) Sandra Sandra F. Zane, EM Tech. Biol. Dept. UNCC Charlotte, NC 28223 sfzane-at-unccvm.uncc.edu Fax (704) 547-3128
Mark, I have had good luck, good quality and quick turn around from Micro Engineering (409)291-6891. My purchases were only resharpennings of old DuPont knives, but the price was good and so was the quality. Some of the folks I work with now will only buy Diatome diamond knives. Hopefully that means that almost any major manufaturer will have some high quality knives available. Doug
Douglas W. Cromey, M.S. Cell Biology and Anatomy Arizona Health Sciences Center 1501 N. Campbell Ave. Tucson, AZ 85724 (520)626-2824 dcromey-at-ccit.arizona.edu
The Electron Microscopy/Imaging Center at Colorado State University will be offering a 5 day Short Course in Freeze-Fracture Techniques this summer. In addition to comprehensive lectures, state-of-the-art instruments will be available for hands-on training in the laboratory.
For more detailed information, please visit our World Wide Web site at URL: http://www.vetmed.colostate.edu/anatomy/emic/homepage.html, or contact me directly (not to this list) for an email copy of the announcement.
John chandler-at-lamar.ColoState.EDU http://www.vetmed.colostate.edu/anatomy/faculty/chandler.html
Mark Elliott asked about diamond knives. Du Pont sold their diamond knife business to Delaware Diamond Knives, Inc. years ago. I don't know the further development. We have several users here; all use Diatome knives and all are satisfied with their knives. One of the users sent a Diatome diamond knife, which had been resharpened by a competitor, back to the Diatome factory for resharpening several years ago. Diatome would not touch it because it was not one of theirs, although the boat was. The diamond was of lower quality. This indicates their high standard of quality. There may be other good diamond knives in the marketplace, but you can't go wrong with a Diatome knife. BTW I have nothing to do with the Diatome except using their knives.
For the past 12 years, I have been dealing exclusively with MicroEngineering Inc. who make the Microstar diamond knife. I have no interest in this company other than the fact that they make a high quality, affordable product. In the rare cases when I have received a bad one, I have had a replacement by the next day or 2. Pricewise, I would have difficulty justifying any other brand to my procurement department. Again, this is no sales pitch, but just the way its been with my lab which buys or resharpens 1 or 2 knives a year.
-=W.L. Steffens=- College of Veterinary Medicine University of Georgia
} From: {MELLIOTT-at-prl.pulmonary.ubc.ca} } Organization: UBC Pulmonary Research Lab } To: microscopy-at-aaem.amc.anl.gov } Date: Mon, 3 Apr 1995 15:18:39 +0800PST } Subject: diamond knives } Priority: normal
} I remember seeing a request in the last few weeks about diamond } knives. We are about to purchase one and are looking for suppliers } and any thoughts on who makes the best diamond knives. I seem to } remember the last time I bought one that the best were from Dupont or } Diatome(?) but can't remember exactly which. Is that still true.? } Any help greatly appreciated. } } Yours } Mark Elliott PhD }
Mark,
I have always used Diatome diamond knives, as did the lab where I was trained. They are of superior quality and can be sharpened as often as needed, for life. Other companies sometimes limit the number of sharpenings that they will guarantee. I also have always had exceptional help and service from Stacey Kirsch who heads the US division of Diatome. She can be reached at 800-523-5874 for current pricing and additional information.
S. sesack-at-bns.pitt.edu Susan R. Sesack Dept. Neuroscience University of Pittsburgh Pittsburgh, PA 15260
CALIFORNIA ASSOCIATION OF CRIMINALISTS MEETING ANNOUNCEMENT
The California Association of Criminalists (CAC) is holding its 85th Semi-Annual meeting on May 10-13, 1995, at the Walnut Creek Marriott in Walnut Creek, California. The Contra Costa County Sheriff's Department Criminalistics Laboratory will be hosting this meeting.
The program is shaping up fast. Some of the subjects planned include: the Integrated Ballistics Identification System (IBIS), computer animation in casework, retrieving secured data in cases of computer crime, and bite mark analysis in the case of a mountain lion attack. This is also our last call for papers so this is your chance to share your latest project or most interesting case. Both your technical notes and innovative research are welcome. For an abstract form contact Rich Schorr, voice (510)646-2455 or fax (510)646-2913.
Workshops planned include: A Computer Workshop for Cyber- phobes (Everything You Ever Wanted To Do But Were Afraid To Admit You Didn't Know How), Practical Applications of GCMS In a Crime Laboratory Environment, a Glock Armorers Course, DNA Users Group Meeting and a Polaroid Film Product Workshop.
For more information or to receive a registration package contact Karen Sheldon, Contra Costa County, Sheriff-Coroner's Department, 1122 Escobar St., Martinez, CA 94553, voice (510) 646-2455 or fax (510)646-2913.
Dear Cryo Electronmicroscopists, I have a student who wants to compare the quantity of extracellular polysaccharide material produced by bacterial cells grown on agar plates (we don't want to use cells from broth) by TEM. The bacteria are involved in plant nodule formation. We would like to loose as little of the material as possible during processing. I think plunge freezing with a KF80, cryosubstituting the cells using perhaps 2% OsO4 in methanol would be good, followed by conventional dehydration and embedding in epoxy. Conventional room temperature fix methods use glutaraldehyde/ruthenium red/OsO4 in various concentrations to fix and stain the extracellular material but from the pictures I've seen I suspect much of the material has still been lost.
Is cryoprotection of the sample necessary? If I lightly fix then infiltrate in sucrose or some other protectant I'm a bit worried we'll start washing the polysaccharide away, if I don't then the cells (which will be a small scraping of colonies off the plate surface) could be ice damaged. If I don't use ruth. red as a stain will the material still be visible - even if present? I'd be grateful to hear from anyone who has had experiance of similar samples.
Regards, Richard E
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
Dear John, You asked about specific improvements in capabilities vs reduced life- times for pointed filaments in use with radiation-sensitive specimens. First, let me say that we experienced no reduction in lifetime. Since, with proper design, a pointed filament can be saturated at a lower current, this is not too surprising. In my work--as opposed to nearly everyone else I know--I often try to get rid of electrons. For EDX and crystallography I operate at the low- est setting for wehnelt bias, a maximally excited first condenser lens, and a small-bore condenser aperture. If I use a more intense beam for EDX, the dead- time of my system and my beam spot both get larger. For crystallography, I scan the specimen using low beam currents to avoid damage during the time I am not recording (damage during recording is inevitable). Furthermore, when I scan in diffraction mode, I want the beam size to be the same as the selected area, so that, again, every electron incident on the specimen is of some use. Bearing these facts in mind, the smaller beam size from a pointed fil- ament reduces the brehmsstrahlung radiation from the condenser aperture and other sources within the column and increases the signal-to-noise ratio and spatial resolution for microanalysis (not that my spatial resolution is too good; with a high-voltage EM and thick sections, the analysis volume will never be small). Exposure of unexamined areas of a specimen is crystallography is harder to characterize; however, given that scanning the specimen, tweaking the position and adjusting the beam, etc. take only a few percent of the expo- sure recorded on film, and that often several ED patterns can be taken from the same selected area, there would seem to be little effect on the data. It must be remembered that the ultimate, irreducible limit to the data obtainable is set by damage to the specimen and that the higher-resolution reflections are the most sensitive to this damage. Once again, it is the smaller size of the beam which is beneficial. Additionally, the ED pattern is better when the ob- jective lens is focussed (although, in the HVEM, this is not too critical), and focussing can be accomplished very rapidly using minimum contrast, which works better with a more coherent beam. I have not done quantitative comparisons between regular and pointed filaments for either case, so I can't give you specific figures of merit, but I can tell you that the beam is visibly smaller and the interference fringes are noticably more prominant with the pointed filament. I also have no idea whether any of these considerations are specific to 1.2 MV electrons, although I wouldn't think so. Another occasional consideration is that heating and charging effects are minimized at low beam currents, so the pointed filaments are more specimen- friendly in this way also. Yours, Bill Tivol
I mentioned in my last message about DIATOME would not touch the knife which had been resharpened by a competitor. I should have made it clear that "the diamond was of lower quality" was a direct quote from a sales rep.
I POSTED THIS MESSAGE SEVERAL WEEKS AGO AND RECEIVED NO RESPONSE. I THOUGHT I'D GIVE IT ANOTHER TRY JUST IN CASE THERE WAS A DELIVERY PROBLEM.
RE: Copy of: Polaron Polabed 812
Dear Microscopists-
I was wondering if anyone out there knows what happened to the Polaron line of products. I am particularly curious about some of their embedding resins such as the Polabed 812.
Thanks for your help!
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
} Dear Cryo Electronmicroscopists, } I have a student who wants to compare the quantity of extracellular } polysaccharide material produced by bacterial cells grown on agar plates } (we don't want to use cells from broth) by TEM. The bacteria are involved } in plant nodule formation. } We would like to loose as little of the material as possible during } processing.
Cryofixation would be a good way to go. Why not freeze sub in osmium/acetone, embed in epoxy, as normal, and then do a PAS stain on sections? The osmium and normal poststains are unlikely to stain all the polysaccharides. It would be interesting to find out if freeze substitution in ruthenium red/acetone would work (i.e., would the RR penetrate?).
R. Howard Berg, Ph.D. Biology Department University of Memphis Memphis, TN, 38152 E mail: bergrh-at-cc.memphis.edu phone: 901-678-4449 fax: 901-678-4457
Continuing the diamond knife theme, I have been using Diatome knives for many years, and have been impressed with their quality and reliabity. But in the commercial exibits at the Cell Biology meetings in San Francisco last December, I became aware of diamond knives being made in the Netherlands by Drukker International, B.V., an old dutch diamond firm (dating from 1906). The descriptions sounded quite impressive, including "superb wetting behavior". A 3 mm blade knife costs $2500. Or you can use their "exchange program" = if you send them an old diamond knife (any manufacturer, any condition), you will receive a new 3 mm Drukker knife for $1750 (if the new knife isn't shipped within two weeks, then you get it free). The U.S. distributor is Harris Diamond Corporation, 100 Stierli Court, suite 106, Mount Arlington, N.J. 07856, tel (201) 770-1520, fax (201)770-1549. I don't know anything about the company other than spending a few minutes at their booth at the meetings and receiving a nice brochure. If I had plenty of money and were buying a diamond knife today, I would probably buy Diatome, but I thought EM people would want to be aware of the Drukker knife, which could be very good.
To listserver people, Many thanks for the replies to my problem. We found that putting the Microcapsules between 2 squares of double sided tape and tearing them apart, left a good idea of the internal structure. Pretty simple really! This was adequate for what the user was wanting and they were pleased with the results. So pleased, they have decided to bring back about 80 samples!
rich.
***************************************************************** * Richard Lander * * South Campus Electron Microscope Unit * * c/- Pathology Department * * Otago Medical School * * P.O. Box 913 * * Dunedin * * N.Z. * * * * Tel. National 03 479 7301 International 64 3 479 7301 * * Fax. National 03 479 7254 International 64 3 479 7254 * *****************************************************************
} From: "W.L. Steffens" {STEFFENS.B-at-calc.vet.uga.edu} } Organization: College of Vet. Med } To: Microscopy-at-aaem.amc.anl.gov } Date sent: Tue, 4 Apr 1995 09:29:07 EST } Subject: diamond knives } Priority: normal
} To Mark Elliot: } } For the past 12 years, I have been dealing exclusively with } MicroEngineering Inc. who make the Microstar diamond knife. I have no } interest in this company other than the fact that they make a high } quality, affordable product. In the rare cases when I have received a bad } one, I have had a replacement by the next day or 2. Pricewise, I would } have difficulty justifying any other brand to my procurement department. } Again, this is no sales pitch, but just the way its been with my lab which } buys or resharpens 1 or 2 knives a year. } } -=W.L. Steffens=- } College of Veterinary Medicine } University of Georgia } MicroEngineering is now known as Micro Star Technologies. Contact: Cathy Zimmerman (800)533-2509, E-mail US3SNQ7N-at-IBMMAIL.COM *************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
Although, its been a while since I've polished aluminum. My initial impression is you have way too many steps. I'd go from 600 grit to 5 micron, to 1 micron, perhaps the .25 micron and then the colloidal. I also remember that cerium oxide paste gave better results. (probably not the greatest idea from a saftey and envirnomental standpoint) THe key to fine polishing of aluminium is to keep the steps to a minimum and the length of time at each step to a minimum. There may be a sure bet way of polishing aluminium....but I found it a real pain.
On Tue, 4 Apr 1995 IE09-at-VM.ACS.UNT.EDU wrote:
} We are currently trying to polish aluminum samples (7075). Before we etch } them, we are trying to get a 0.05 micron finish. The grinding/polishing } sequence is: 600 grit, 800 grit, 1200 grit, 3 micron diamond paste, 1 micron } diamond paste, 0.25 micron paste, 0.05 colloidal silica. Everytime we go from } the 1 micron step to the 0.25 micron step scratches appear.... } } We have been using Allied High Tech Spec-Cloth polishing pads. Doeas anyone } have suggestions to get to the final two steps without introducing scratches? } } I do not really understand why they get scratched so easily when the finer } pastes are used. } } Should these samples be stored in a dessicator to prevent oxidation? I have } never worked with Al, and therefore would appreciate any suggestions. } } Patrick Diehl } Center for Materials Characterization } University of North Texas
} Date: 05 Apr 95 12:18:19 EDT } From: South Bay Technology {73531.1344-at-compuserve.com} } To: Microscopy ListServer {MICROSCOPY-at-aaem.amc.anl.gov} } Subject: Copy of: Polaron Polabed 812
} I POSTED THIS MESSAGE SEVERAL WEEKS AGO AND RECEIVED NO RESPONSE. I THOUGHT I'D } GIVE IT ANOTHER TRY JUST IN CASE THERE WAS A DELIVERY PROBLEM. } } RE: Copy of: Polaron Polabed 812 } } Dear Microscopists- } } I was wondering if anyone out there knows what happened to the Polaron line of } products. I am particularly curious about some of their embedding resins such } as the Polabed 812. } } Thanks for your help! } } David Henriks } South Bay Technology, Inc. } 1120 Via Callejon } San Clemente, CA 92673 } } TEL: 714-492-2600 } FAX: 714-492-1499
David
I am not sure about specific products but the fate of Polaron goes Polaron was sold to BioRad who sold it to Fisons. By contacting a US rep of Fisons you may get more info, although the unit may have been sold to Thermo Instruments as part of the Fisons scientific instrument division sale.
As far as Polabed goes a number of suppliers have similar alternative to EPON 812.
Ian Hallett
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660
} } We (DuPont CR&D Analytical) have hosted girls in our lab for the past 2 years } on Take Your Daughters to Work Day and have set up several demonstrations using } materials we thought would be familiar to the girls. Lots of parents bend the } rules and bring quite small girls as young as 6, so be prepared. } My High School teacher wife read this over my shoulder and asked me to remind everyone who has these types of event to have them in the summer so that the students won't miss school. Teachers who try to do a good job for classrooms of 25 to 35 students, cannot tailor their lessons to meet the demands of students who take days off for going to work with mom or dad, and similar things can can just as easily be done during school vacation periods.
It is difficult to pinpoint the exact cause for the scratches, but there are a few things you may want to check.
1) What size are the scratches that you see? Are they significantly larger than the 1u scratches you are trying to remove? Perhaps you are getting an agglomeration of .25u particles if they are not properly wetted and dispersed in the paste. Have you used this same .25u paste before without trouble? Perhaps you should try a diamond suspension that has more uniformly dispersed particles.
2) Is your sample mounted in epoxy? If so, perhaps particles are emedding in the epoxy from a previous polishing step.
3) If your sample is not embedded in epoxy, perhaps the particles are embedded in the mounting wax or trapped inside the polishing fixture.
I don't think your cloth would be a problem. What you are using is the same as our Fine Rayon cloth and that is what I would recommend for what you are doing. I assume that you are using a fresh cloth when you reach this stage. Of course, oxidation could be the problem, but I have never had that problem so it doesn't seem likely to me.
Something else you may want to consider is trying Colloidal Alumina for the final polishing stage instead of Colloidal Silica. I understand that it can produce a better finish on softer metals. Of course, I understand you are having the problem before the colloidal silica, but it may be something to consider when you get to that point.
Of course, I would be pleased to supply you with a sample of the diamond suspension or the colloidal alumina if you would like to give that a try. I would also be interested to hear any suggestions you get from other people and ultimately your determination of the problem. I wish you good luck in figuring it out and I encourage you to contact me if I can be of any help.
Best regards-
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
Further continuation of the diamond knife theme::
1] I have heard essentially good things about the Drukker company. The only negatives I have ever heard came from Drukker competitors.
2] The Drukker people at the ICEM 94 in Paris and at MSA in 94 had in their exhibit both a video demonstration showing some purported advantages of their knife, because of the hydrophilic nature of the knife, over other knives. The advantage supposedly was that the sections come off more smoothly and with less vibration, e.g. slipping and sliding down the edge of the blade. At the ICEM 94 in Paris, the Drukker booth attracted some of the largest crowds.
Can anyone comment as to whether they have actually seen the hydrophilic edge to be a unique advantage, above and beyond diamond knives generally and therefore worth spending extra money?
Further with regard to Drukker, it is my understanding that they more or less "own" the world wide business for diamond scalpel blades used for radial keratotomy procedures being done by ophthamologists. They are now trying to apply their expertise gained from the surgical instruments end of the business to ultramicrotomy.
3] The "talk" seems to center around foreign made diamond knives. There are some excellent US made knives (e.g. MicroStar and DDK) and again, standing in our exhibit booths at the various meetings, I don't detect any unhappiness among their customers either. As a group, they do seem to be "happy campers". But as the US dollar stays weak relative to European currencies, those campers are going to have another reason to be "happy": They are going to be saving money.
Charles A. Garber PRESIDENT SPI SUPPLIES/STRUCTURE PROBE, INC. PO BOX 656 West Chester, PA 19380 USA
} Dear Cryo Electronmicroscopists, } I have a student who wants to compare the quantity of extracellular } polysaccharide material produced by bacterial cells grown on agar plates } (we don't want to use cells from broth) by TEM. The bacteria are involved } in plant nodule formation. } We would like to loose as little of the material as possible during } processing. I think plunge freezing with a KF80, cryosubstituting the cells } using perhaps 2% OsO4 in methanol would be good, followed by conventional } dehydration and embedding in epoxy. Conventional room temperature fix } methods use glutaraldehyde/ruthenium red/OsO4 in various concentrations to } fix and stain the extracellular material but from the pictures I've seen I } suspect much of the material has still been lost. } } Is cryoprotection of the sample necessary? If I lightly fix then infiltrate } in sucrose or some other protectant I'm a bit worried we'll start washing } the polysaccharide away, if I don't then the cells (which will be a small } scraping of colonies off the plate surface) could be ice damaged. } If I don't use ruth. red as a stain will the material still be visible - } even if present? } I'd be grateful to hear from anyone who has had experiance of similar samples. } } Regards, } Richard E } } Richard Easingwood } South Campus Electron Microscope Unit } Otago Medical School } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } "The southernmost electron microscope unit in the world"
You can try the method devoloped in Mueller's Lab, ETH, Switerland by using a capillary tube. "High-pressure freezing of cell suspensions in cellulose capillary tubes", J. Microscopy 175, 34-43 (1994).
Ya Chen
========================================================================= \ / Assistant Researcher-Cryo/SEM Coordinator TEL : 608-263-8481 \ / __ Integrated Microscopy Resource (IMR) FAX : 608-265-4076 \/ / / University of Wisconsin-Madison / / / 1675 Observatory Drive #167 Email:YChen-at-macc.wisc.edu / /__/_ Madison, WI 53706 Email:chen-at-calshp.cals.wisc.edu =========================================================================
I am not familiar with the Allied Spec-Cloth polishing cloths you are using, nor have I polished 7075 Al (to my knowledge), but here is some suggestions for ya to do with as ya see fit:
Using Buehler products, grind on grits 320, 400, 600, and 800; polish on: 1. 6 micron diamond paste [water-based] on Texmet, 2. 1 micron diamond paste [water-based] on either Texmet or Metcloth, 3. either (a) 1 micron Cerium Oxide solution on Mastercloth -or- (b) .5 micron Chrome Oxide solution on Mastercloth, then 4. .05 micron Mastermet [colloidal Silica] on Mastercloth.
Few hints: - make sure new cloths are used, and that before their use, you "beat" down the fibers on the Mastercloth by using a dummy/blank sample or such; - if using a ultrasonic to help in cleaning, this may dislodge particles that will then add scratches in later steps; - use very warm water and -much- soap to clean the samples between each step. I use my thumb to very gently rub and clean the sample surface; - use of a dessicator is highly recommened to help slow oxidation and to keep samples cleaner for further examination; - takes much time and is -very- frustrating, but experiment with the above and other recipes to get what works best for you; - have not done much myself, but don't overlook electropolishing if such can be used in your case; - for me, water-based diamond pastes seem to "cut" better for rough polishing, whereas oil-based seem to produce a smoother, cleaner finish; - if get frustrated, quit if can for awhile, and attack the sample(s) later on - have found myself that a recipe that works wonders one day seems to suck later, but then seems to work great again! The joys of hand-polishing!! oh boy...
Hope this is somewhat helpful. Good luck, -Rob disclaimer: [once again] am not affiliated nor have any financial interest in any products mentioned or used ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /\v/\ | | Metallographic Lab. | Missouri Speleological Survey /\v/\ | | Rolla Research Center | Bat Conservation International /\v/\ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-ptbma.usbm.gov | National Speleological Society #32993 /\v/\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
} You can try the method devoloped in Mueller's Lab, ETH, Switerland by using } a capillary tube. } "High-pressure freezing of cell suspensions in cellulose capillary tubes", } J. Microscopy 175, 34-43 (1994). } } Ya Chen } Does anyone know of a US Supplier of dialysis tubing in the dimensions specified in Mueller's paper (about the diameter of a capillary tube)?
R. Howard Berg, Ph.D. Biology Department University of Memphis Memphis, TN, 38152 E mail: bergrh-at-cc.memphis.edu phone: 901-678-4449 fax: 901-678-4457