owenha-at-csd.uwm.edu (Heather A Owen) Message-ID: {1996Oct01.101559.1814.79639-at-missgate.sunderland.ac.uk} X-Mailer: Microsoft Mail via PostalUnion/SMTP for Windows NT Mime-Version: 1.0 Content-Type: text/plain; charset="US-ASCII" Organization: University of Sunderland
Heather
my knowledge of the workings of our LKB multiplate are extremely limited but I would suspect, if you are getting heat but not enough, that there may be a faulty thermostat rather than heater. Sorry I don't know where to get one of those either.
Malcolm Haswell E.M. Unit University of Sunderland U.K.
----------
Does anyone know of a vendor for a replacement heater for an LKB Multiplate apparatus that heats wax for sealing boats to glass knives. I put a thermometer into the one we have, and it only reads 62 C in the wells and the same on the knife warming deck (the instructions indicate that the temperature should be about 80 C). The wax (pink dental) is barely melted, and we have a very short window of opportunity before the wax solidifies. I'd appreciate any information.
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Message-Id: {1.5.4.32.19961001131004.0066b690-at-mailhost.ultra.net.au} X-Sender: pns-at-mailhost.ultra.net.au X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 20:00 30/09/96 -0400, you wrote: } I have recently been using a Hitachi S-570 SEM. I wanted to use Polaroid 53, } but this being an older machine it does not have a setting for 800 speed } film. I was considering using the recommended setting for 400 ISO film and } close the lens aperture down one stop. Does anyone know if this is feasible } or can they recommend a better method. } } Thank you } Tom Bryner ****************************** Hi Tom and whoever: I believe that you cannot do better than to close down the camera by one stop. The only other consideration is lens resolution. For that size format best resolution for most lenses would be between f5.6 to 11. Lenses designed for 35mm format are likely to have best resolution with the aperture one stop more open. If the image is properly in focous, the additional depths of field is no consideration when photgraphing a flat screen. Cheers Jim Darley Probing & Structure } (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } A great microscopy site http://www.ultra.net.au/~pns/
A belated thank you to the few of you who replied to my posting about persistent white atoms in HRTEM simulations. It would appear that Mike O'Keefe hit the nail squarely, since the white atoms disappear when I add 16 milliradians of crystal tilt to the simulation. They are also not present for very small specimen thicknesses.
Also, thanks to Larry Allard I will spell "Scherzer" correctly for the rest of my life.....
Roy Christoffersen Texas Center for Superconductivity 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
Should be. We do it often with our JEOL 840. We normally run 100 seconds at f-5.6 for Type 55 film. If I am in a hurry, I will shorten the exposure to 50 seconds and open the aperture to f-4. Each f-stop change will double or halve the light coming through, so just expose it half or twice as long, respectively.
At 08:00 PM 9/30/96 -0400, you wrote: } I have recently been using a Hitachi S-570 SEM. I wanted to use Polaroid 53, } but this being an older machine it does not have a setting for 800 speed } film. I was considering using the recommended setting for 400 ISO film and } close the lens aperture down one stop. Does anyone know if this is feasible } or can they recommend a better method. } } Thank you } Tom Bryner ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
Hello Tom, Yours is an excellent solution since one larger f-stop number (not larger opening) is about the same as one/half the light or twice the film speed. Besides, your depth of field and therefore , focus may be better.
Good luck. Alex Greene Scientific Instrumentation Services, Inc. [Independent Electon Microscope Service] Number 499, Post Office Box 19400 Austin, Texas 78760
At 08:00 PM 9/30/96 -0400, you wrote: } I have recently been using a Hitachi S-570 SEM. I wanted to use Polaroid 53, } but this being an older machine it does not have a setting for 800 speed } film. I was considering using the recommended setting for 400 ISO film and } close the lens aperture down one stop. Does anyone know if this is feasible } or can they recommend a better method. } } Thank you } Tom Bryner } } }
Hi All, we have been trying to find different ways to prepare SOI specimens for TEM. The material of interest is a silicon wafer which has an oxide lare on it, on top of which there is the silicon (SOI) layer. This layer can be as thin as 0.1-0.2microns, and the oxide can be 0.1-0.8 microns. It would be nice if we could lift off the top layer only as a TEM specimen. I have tried placing in an HF bath, however it takes sufficiently long to get thorough all the oxide, that you start seeing hjoles in the top layer too. I was trying to figure out a way to coat the top layer with something robust and non-damaging and inert (or reacts slowly with HF), and which could be easily removed after lift-off. I thought of super-glue or wax, but thought I'd ask if anyone has had similar experiences before I try it out.
Any suggestions?
cheers
Lucio
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr.Lucio Mule'Stagno MEMC Electronic Materials Inc University of Missouri -St.Louis Silicon Materials Research Group Physics Dept., 501 Pearl Dr., 8001 Natural Bridge rd., St.Peters, St.Louis MO 63376 MO 63121 tel: 314 279 5338 314 516 5931/3 fax: 314 279 5363 314 516 6152 email: lmulestagno-at-memc.com luciom-at-newton.umsl.edu URL:http://www.newton.umsl.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
To: Tom Bryner {brynert-at-mcmaster.ca} Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com} *** Reply to note of 10/01/96 08:51
Tom,
What you suggest for setup is fine, but you need to go further.
Most SEM's have a "graph" mode that shows the video signal intensity across a line on the image. You need to mark the CRT bezel (or similar reference) for where the image signal is barely white on the top, and barely black on the bottom.
A high contrast sample with full grey scale range works nicely. Trial and error until you get an OK photo, and then note areas where you just barely maintained highlight detail and one where you just barely maintained shadow detail. Look at the graph in these areas and mark the levels on the bezel. This has to all be done with no changes in the setup, and the beam must be stable. If your getting level drifts, you'll have to settle it down 1st. After that, you're set with very exact repeatable reference marks to go by. Try it, and let me know how you do.
I was asked by our very recently appointed User's Committee of the EM core facility of the university which has been given the task "to explore options for the future of the lab" to find out how other similar university facilities are supported. In particular, how much support is provided by the central administration, the colleges and/ or departments for the running of the lab? Also, where do such labs usually sit in the infrastructure of the university, ie. in a college,underthe vp for research or other entity? The university,I guess is a midsize institution of around 15,000 so I believe a core type facility makes sense in reducing redundancy, costs, etc., but so far the users and myself are having a hard time convincing the central administration this. Thanks in advance Hank Adams EML New Mexico State University
Closing the aperture by one stop is the simplest solution.
Another method you might consider (about which I'd welcome comments from the EM community) is to recalibrate the output levels of the record screen amplifier to match the faster film. The benefits are;
1) you will know for sure that the output matches the film rather than relying on "what should be the case" (and possibly being thwarted by unforseen irregularities in the system). Also, I gather that the phosphor in the screens "ages", so if it's been a long while since you've matched output to film, you should probably do this anyway.
2) The width of the scan line of the record screen is related to the brightness. I assume this is due to bright phosphor particles exciting neighbouring particles. Excessive brightness could ultimately, I expect, produce overlapping scan lines, reducing the effective resolution of the record screen image - this appeared to be the case in our SEM. (Note: you can also lose resolution if the electron beam in the CRT is not properly focussed - this is adjustable on our SEM). To cut a long story short, I opted to match the film-speed to my chosen output level of the record screen (maintaining optimal f-stop) rather than the normal way of matching output to film/camera.
In your case, retuning the output to a faster film would entail reducing the brightness. This would be going in the right direction for reducing the variation in width of the scan line on the record CRT.
I'd welcome comments on this from others, particularly if it introduces other detrimental effects of which I'm not currently aware.
Cheers, Geoff
:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: Geoff Avern Manager Microscopy Laboratories Australian Museum Email: geoffa-at-amsg.austmus.oz.au 6 College St Ph: (61)(2) 9320 6198 Sydney, Australia. 2000 Fax: (61)(2) 9320 6059 ::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
I have recently been using a Hitachi S-570 SEM. I wanted to use Polaroid 53, but this being an older machine it does not have a setting for 800 speed film. I was considering using the recommended setting for 400 ISO film and close the lens aperture down one stop. Does anyone know if this is feasible or can they recommend a better method.
On a recent visit to Hitachi Naka Works I was shown a Focussed Ion Beam Miller (the FB2000). They are commonly used in semiconductor defect analysis. Does any one know who else makes them?
Dear Tom, If you can stand another message on your S-570, here's mine. I also have an S-570. Yes, closing the f-stop one stop should adjust the camera for the 800 ASA film. Also, if you need to adjust the camera slightly, there are adjustments for photo CRT brightness and contrast on the panel just to the left of the camera, under the cover. If you run out of adjustment, there are coarser adjustments in the back that the Hitachi service people can tweak. Hope this helps, Regards, Mary
Tom Bryner wrote: } I have recently been using a Hitachi S-570 SEM. I wanted to use Polaroid 53, } but this being an older machine it does not have a setting for 800 speed } film. I was considering using the recommended setting for 400 ISO film and } close the lens aperture down one stop. Does anyone know if this is feasible } or can they recommend a better method. } } Thank you } Tom Bryner
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Dear Eugene: The definition of "video" is the 640 X 480 size. You must use a higher resolution camera to get a higher resolution image. These, however, will not be compatible with a video VHS recorder and will not work at real time speeds.
Eugene Krueger wrote: } The lab I'm in uses time lapse recorders (Super VHS) for videoing live cells } on a microscope stage under different conditions. The resulting videotape } looks terrific on a TV/ monitor. The problem is when I videocapture from the } tape (for posters or papers), the resulting image is limited to 640 x 480 } pixels even if I set my monitor as high as 1024 x 768. Is this a limitation } of the software/hardware set-up I'm using, or is this the limit of resolution } of VHS/ SVHS tapes? I perform the captures on a PowerMac 8500 using a SVHS } recorder and the built-in SVHS port of the mac. The capture software is Apple } Video Player (free with the mac). Would a commercial software product allow } me to capture "larger" images? Would a different videoboard matter? } Luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Message-Id: {1.5.4.32.19961002042609.0066c368-at-mailhost.ultra.net.au} X-Sender: pns-at-mailhost.ultra.net.au X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 18:24 30/09/96 -0500, you wrote: } } Does anyone know of a vendor for a replacement heater for an LKB } Multiplate apparatus that heats wax for sealing boats to glass knives. I } put a thermometer into the one we have, and it only reads 62 C in the } wells and the same on the knife warming deck (the instructions indicate } that the temperature should be about 80 C). The wax (pink dental) is } barely melted, and we have a very short window of opportunity before the } wax solidifies. I'd appreciate any information. } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816 ************************************************* Hello Heather and other multiplaters: LKB was swallowed up by Leica and they should have spare parts. The gadget does not have a thermostat but relies on the heater to reach a design maximum which is a bit over 70 degrees. This could be limited by low ambient temperature, poor heat transfer from the element and wrong operating voltage. If the element is designed for 120 volts but the line voltage is only 110, considerably lower temperatures will result. Your electronic technicians will have some heat transfer paste. Smearing some of that over the cylinder shaped element may do the trick. I prefer a small adjustable lab hot plate with an Al gadget on top to hold wax and spatula. I prefer to seal glass boats with dental wax rather than nail varnish; I think that poor temperature control of the dental wax is the main reason that varnish is used. Cheers Jim Darley Probing & Structure } (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } A great microscopy site http://www.ultra.net.au/~pns/
Dear Glenn, The best reference I know is Bastin et al in the MAS Conference Proceedings of five or six years ago. He did a very thorough study of light element analysis spanning several years and including a study of contamination, since it is so important in light element analysis. He also attempted to work out a method of analysing the C in steel by graphing the apparent C level and extrapolating back to time = 0. He also found that most contamination is specimen derived. I attempted some C in steel several years ago and on "good" days it was almost possible. Use low kV and as low a current as you can to get the x-rays. I then attempted to study the contamination rates on a piece of polished oxygen-free copper and found that it varied widely and without any logical reason. One source I found was if the e-beam hit a bit of mounting epoxy, the contamination level was high for three days after. I attempted to use an air-jet that injected air or oxygen on to the sample to remove contamination or prevent its deposit, but it only worked once in five days of trying, and I don't know why. One thing I tried for the C in steel was to put the sample into the camber the evening before the analysis, so it pumped overnight and was not vented in the morning before the analysis was done. It is always a problem. Hope this helps Regards, Mary
Glenn Poirier wrote: } Does anyone know of a reference for information on carbon contamination } rates in microanalysis (i.e. effect of current, accelerating voltage, } spot size etc. )? I'm going to try to do some semi-quantitative } analyses of carbon in steel and am trying to find analytical conditions } which minimize contamination. Also, if anyone knows of a technique(s) for } cleaning samples before carbon analysis could they share it with me. } } TIA } } Glenn
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Could anybody out there help me? I am looking for an imaging video system which can digitalize what is observed in a microscope, and send to a screen (computer of TV set). I will really appreciate your opinions about the best ones.
Thank you in advance
------------------------- Carlos A. Canchaya Genetic Resources and Plant Biotechnology Research Center National Agrarian University Lima-Peru inquil-at-amauta.rcp.net.pe
In addition to the remarks by Mary Mager: G. F. Bastian and H. J. M. Heijligers from Laboratory for Physical Chemistry, University of Technology, Eindhoven, Netherlands have written a detailed report on the matter. The title is "Quantitative electron probe microanalysis of carbon in binary carbides" and its ISBN number is 90-6819-004-0 CIP.
Regards, Joergen. -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--at--at--at- J. B. Bilde-Soerensen Materials Department Risoe National Laboratory DK-4000 Roskilde Denmark
I would like to contact with people who knows or is using the Universal Microscope of Aus Jenna. We have one in our lab and it have most of his parts and accesories but we do not know how to assemble it. There is no representant of Aus Jenna in my country. We do not also have its model and date of fabrication. It seems that it is a very good microscope for research. My questions are:
- Does anybody know how to obtain its handbook or manual of its parts and how to use it? - Do you know information about what the model is and when it was fabricated? - Does Aus Jenna have a Web site in Internet? - Could anyone suggest the best alternative for that microscope, does the alternative have all the same features of the Universal (fluorescence, phase-constrast,inverted microscopy etc)
We really appreciate your future help.
Thank you in advance,
------------------------- Carlos A. Canchaya Genetic Resources and Plant Biotechnology Research Center National Agrarian University Lima-Peru inquil-at-amauta.rcp.net.pe
Does anyone knows some Beem flat embedding molds from polyethylene deeper than 3mm? we have some problems with embedding media such as nanoplast, lowicryls or LR-white...because the bloc is too thin and sometimes it breaks. Thanks in advanced Nuria Cortadellas
Thanks for answer my question refered to the tensile and torsion test by SEM, what you send me, was an important help to complete my thesis, thanks again
If you have access to the world wide web, a great place to start it the NIH Image home page: http://rsb.info.nih.gov/nih-image/ NIH Image is image processing, acquisition share-ware that works on the Mac. There will soon be Windows (95?) version as well. There are thousands of users around the world and many derivatives of the program and wide commercial support. There is also a very active NIH Image listserver similar to this one.
We use several frame grabber boards. My favorites: Data Translation Quick Capture for Mac (from video source) , and Data Translation 3152 for PCI-bus Win 3.1(from analog & digital SEM signals) We have all but eliminated the use of Polaroid film in our lab with savings of many thousands of dollars (US) per year.
------------------------------------------------- Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-rd.sylvania.com
Our web sites: www.sylvania.com www.osram.de www.siemens.com
I'm looking for 5 .tif file SEM images of common objects for a contest at the Prince George Science and Technology week. These micrographs will be published in the paper 1 at a time for 5 days and then the kids will guess what they are and a prize will be given to those with the most correct.
I volunteered to give the Sci and Tech week committee these micrographs. Our machine was down at the time and now I've found out that the repair parts will not be in until after the deadline so I can't produce them. I know that there are lots of image pages on the web. However, since I need free micrographs that can be published in the local paper I thought asking in this way would be preferable.
Thank you very much for helping me out. Please attach files to e-mail as jpg or tif and e-mail me directly. There is no money for any of us in this, it's just for fun and education.
Jorge Canchaya wrote: } } Hi: } } Could anybody out there help me? I am looking for an imaging video } system which can digitalize what is observed in a microscope, and send } to a screen (computer of TV set). I will really appreciate your opinions } about the best ones. } } Thank you in advance } } ------------------------- } Carlos A. Canchaya } Genetic Resources and Plant Biotechnology Research Center } National Agrarian University } Lima-Peru } inquil-at-amauta.rcp.net.pe
Hi, It depends on what you want to look at. If you are interested in histopathology (as I do) and make image archiving and analysis, you would need a high resolution color camera (3CCD or, better, a digital camera). for the CCD technology you would need a framegrabber (Matrox has a wide range of offer), for the digital camera you need a SCSI port on the PC. the final output will be an image file that is small or huge (for usual CCD cameras and framegrabbers is 1-2MB, for the digital camera you usually have much bigger files). An important item is the PC: try to get a Pentium PC (PCI structure) or a PowerMac. Then you can download the image analysis software from NIH for free or buy a more sophisticated program. Another important thing is the video monitor: I think the best is a SONY Trinitron. If you are interested in time-lapsed live video microscopy a B/W CCD camera is sufficient and the cost is more affordable, but you may need a more sophisticated hardware for image storage and retrieve (i.e. speedy optical disks and software for movie reconstruction). In conclusion you may need to pay anything between 8,000 and 60,000 USD for a decent system. Is up to you.
best wishes
Paolo Ghiara Dept of Immunology - Chiron Biocine SpA, Italy ghiara-at-sienanet.it
} At 20:00 30/09/96 -0400, you wrote: } } I have recently been using a Hitachi S-570 SEM. I wanted to use Polaroid 53, } } but this being an older machine it does not have a setting for 800 speed } } film. I was considering using the recommended setting for 400 ISO film and } } close the lens aperture down one stop. Does anyone know if this is feasible } } or can they recommend a better method. } } } } Thank you } } Tom Bryner } ****************************** } Hi Tom and whoever: } I believe that you cannot do better than to close down the camera } by one stop. The only other consideration is lens resolution. For that size } format best resolution for most lenses would be between f5.6 to 11. Lenses } designed for 35mm format are likely to have best resolution with the } aperture one stop more open. } If the image is properly in focous, the additional depths of field } is no consideration when photgraphing a flat screen. } Cheers } Jim Darley } Probing & Structure } } (Microscopy Supplies & Accessories) } PO Box 111, Thuringowa QLD 4817 Australia } } } } Phone +61 77 740 370 Fax: +61 77 892 313 } } A great microscopy site http://www.ultra.net.au/~pns/ } Years ago I did a study on the appropriate F stop to set the camera on the image recording unit for the SEM (JEOL-35). That instrument had a F4.5 lens and the ability to adjust the contrast and brightness independently.
The end result was interesting. The quality parameter measured was the number of line pairs resolved on the photographic image (Polaroid type 55 was used since it has the resolution to see 2000 line pairs over the 4X5" image. The best result was F8. At smaller F stopsl (F11 and lower) the brigtness had to be increased which had the result of blooming- the phosphor was being pushed too hard. At larger F stops there was a lack of depth of field field resulting in the outer corners of the image being out of focus. Also noticed was defocusing of the image due to "mottling" of the negative material.
Incidentally, our MRS-3 standard has patterns to help check CRT resolution.
Hope this little tutorial helps.
Joe Geller Geller Microanalytical Lab Topsfield, MA 01983 www.gellermicro.com
I cannot help you in finding the data you are looking for, but I must comment some facts. The microscope in question is obiously German made, the city where the factory is located is JENA (not Jenna), and "aus" means "from" in German, so you should look at this as "an universal microscope from Jena". By the way, Jena is the home city of Carl Zeiss Jena, geographically located in the Eastern part of Germany. Hope someone can help you more in this matter, and maybe if you are looking either for "Zeiss" or for "Jena" on the Web you might find something useful. Good luck! Kris
************************************************************************* Kristof KOVACS Associate Professor University of Veszprem, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684 *************************************************************************
Hi All, we have been trying to find different ways to prepare SOI specimens for TEM. The material of interest is a silicon wafer which has an oxide lare on it, on top of which there is the silicon (SOI) layer. This layer can be as thin as 0.1-0.2microns, and the oxide can be 0.1-0.8 microns. It would be nice if we could lift off the top layer only as a TEM specimen. I have tried placing in an HF bath, however it takes sufficiently long to get thorough all the oxide, that you start seeing hjoles in the top layer too. I was trying to figure out a way to coat the top layer with something robust and non-damaging and inert (or reacts slowly with HF), and which could be easily removed after lift-off. I thought of super-glue or wax, but thought I'd ask if anyone has had similar experiences before I try it out.
Any suggestions?
cheers
Lucio
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr.Lucio Mule'Stagno MEMC Electronic Materials Inc University of Missouri -St.Louis Silicon Materials Research Group Physics Dept., 501 Pearl Dr., 8001 Natural Bridge rd., St.Peters, St.Louis MO 63376 MO 63121 tel: 314 279 5338 314 516 5931/3 fax: 314 279 5363 314 516 6152 email: lmulestagno-at-memc.com luciom-at-newton.umsl.edu URL:http://www.newton.umsl.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I would like to contact with people who knows or is using the Universal Microscope of Aus Jenna. We have one in our lab and it have most of his parts and accesories but we do not know how to assemble it. There is no representant of Aus Jenna in my country. We do not also have its model and date of fabrication. It seems that it is a very good microscope for research. My questions are:
- Does anybody know how to obtain its handbook or manual of its parts and how to use it? - Do you know information about what the model is and when it was fabricated? - Does Aus Jenna have a Web site in Internet? - Could anyone suggest the best alternative for that microscope, does the alternative have all the same features of the Universal (fluorescence, phase-constrast,inverted microscopy etc)
We really appreciate your future help.
Thank you in advance,
------------------------- Carlos A. Canchaya Genetic Resources and Plant Biotechnology Research Center National Agrarian University Lima-Peru inquil-at-amauta.rcp.net.pe
BIOMET:Applied Phycological Research and biological microscopy consultancy.
I am looking for some help. I am trying to dissect out some very small {50 um blood vessels. The main problem is that I cannot see them once the blood has washed out. I would like to stain the vessels with something that is compatible with immunohistochemical procedures using confocal microscopy. I am concerned that the vessel staining will interfere with the fluorescence when viewed on the confocal either by scattering or autofluorescence. Since alkaline phosphatase is present in most vessels and is still active after paraformaldehyde fixation, this should would work for the vessel stain. I am using rats and most likely the method of Mayahara et al., Histochemie 11:88;1967. Any advice would be appreciated.
In regards to my previous request for information regarding a non-profit lab c ontemplating going into competitiion with commercial labs, I would like to thank those that responded to me with their comments. Here are some additional details that answer your questions.
The location should not matter. They want to do it to survive so that they can get their own work done. They'll probably only need about 25% from outside sources, so it is believed....They would not turn a profit since any work would be done at cost. Their current equipment inventory does not include any federally funded instruments. We cannot find any laws that address the unfair competition possibility. They do not see it any different than a bookstore at a school being in competition with commercial bookstores. The activity has not yet started but is likely to unless we find some legal objections.
Any further input would be appreciated.
Anne Esposito E.M.C., e-mail: Silverstf-at-aol.com
G'day from Oz, Please would any of you wonderful microscopists out there have looked at Penguin Eyeballs under LM or EM. I'm particularly interested in the retina structure under SEM or TEM. Thank you Gerry
Ms Geraldine Nash Electron Microscopist
EM Unit Australian Antarctic Division Channel Highway Kingston Tasmania 7050 Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351
FEI Company 7451 NE Evergreen Parkway Hillsboro, OR 97124-5830 USA voice: (503) 640-7500 fax: (503) 640-7509
Good Contacts there would be Phil Fischer and Brent Townsend. There are also European contacts: FEI Europe Ltd. Cambridge, UK 44-0954-250-526
FEI Europe, GmbH, Munich, Germany 49-894-623-450
A recent FEI technical newsletter ("FEI focus") says that they'll be at the following trade shows through the end of this year: American Vacuum Soc., Oct. 15-17, Philadelphia, Pennsylvania, US Semicon/Southwest, Oct 15-16, Austin, Texas, US ISTFA '96, Nov 18-20, Los Angeles, California, US MRS, December 2-5, Boston, Massachusetts, US Semicon/Japan, Dec. 4-6, Makuhari, Japan
If the machine that Mel Dickson saw was a "FIB 200," it was an FEI model.
FEI also makes systems ("DualBeam") that combine an FIB system with an SEM.
} } } Scott D. Walck WL/MLBT {walcksd-at-ml.wpafb.af.mil} 10/02/96 06:44 } } } } On a recent visit to Hitachi Naka Works I was shown a Focussed Ion Beam } Miller (the FB2000). They are commonly used in semiconductor defect } analysis. Does any one know who else makes them? } } Thanks, } } Mel Dickson } E.M. Unit, } UNSW, Sydney, Australia } There's two in USA:
Micrion 1 Corporation Way Peabody, MA, 01960 USA (508) 531-6464
FEI 19500 NW Gibbs Dr., Suite 100 Beaverton. OR 97006 USA (503)690-1500
For your information, there will be two talks on FIB sample preparation at the TEM Sample Preparation Symposium Z at the Spring '97 Materials Research Society meeting in San Francisco.
We've just started capturing images directly to the powermac using the built in frame grabber, and not a problem - you can set the resolution to whatever the computer is capable of - for single images, we find a canvas size of 1024x768 and millions of colours is fine, and so far have used only the built-in maximum size for PAL or NTSC when capturing video. You do need mega amounts of RAM though, and need to use at least thousands of colours if capturing colour images. We've used both Apple video player and Avid Videoshop, both of which came with our machine. Maybe if the S-VHS recorder is limiting the resolution, this would be a way to go. It shouldn't be too hard to set up a time lapse system controlled by the computer.
cheers,
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Clayton, Victoria 3168, Australia phone 61-3-9905 5670 email r.g.white-at-sci.monash.edu.au fax 61-3-9905 5613 or rgwhite-at-vaxc.cc.monash.edu.au
I'm a high school teacher whose been given a vintage 1980 Cambridge StereoScan 250 SEM. It is up and running as well as expected but now I need to figure out how to get it more flexible. The recording device is a Polariod affair that is much too expensive for numerous high schoolers. What was intimated was hooking up a digitalizer to the TV mode of the viewing screen and bypass the HRRT.(?) It was also told to me that if we wanted to used the HRRT to capture the image digitally that it would take a bit more money as well as smarts. (Two things I'm rather short on right now).
My mission (of which I am pleading for help with) is to find a product and a vendor of such a digitalizer, a price, and all the pertinent information to present at a meeting next week.
Carl Zeiss of old West Germany had merged with Jena which was the old Carl Zeiss in East Germany before WW2. Zeiss should have a web page in any of the browsers such as Yahoo or try www.zeiss.com and see what happens.
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Nuria Cortadellas wrote:-
} Does anyone knows some Beem flat embedding molds from polyethylene deeper } than 3mm? we have some problems with embedding media such as nanoplast, } lowicryls or LR-white...because the bloc is too thin and sometimes it } breaks. } Thanks in advanced } Nuria Cortadellas
TAAB in the UK make an embedding mold that is similar to a standard type capsule except that it has a full flat end. They are 8mm in diameter. They have a snap lid which is helpful if you are embedding resins like Lowicryl and LR White where you may wish to exclude air. Orientation of specimens in the end of the capsule can be difficult but is usually possible with a bit of ingenuity.
I don't know if TAAB products are available in your locality from another EM supplier. If not then details are:-
TAAB Laboratories Equipment Ltd. 3 Minerva House, Calleva Industrial Park Aldermaston, Berkshire, England. RG7 8NA Phone 01734 817775 (local UK number) Fax 01734 817881 (local UK Number)
I was in Jena (in former East Germany) about two years ago. The entire Jena Zeiss complex was demolished for redevelopment as (I beliece) a shopping complex. Only the original planetarium was remaining, at a separate location in the city.
The Jena company reunited with the Oberkochen company a little while ago. There is a web site at:
http://www.zeiss.de
This also gives phone and fax for Carl Jeiss Jena GmbH: tel. ++49 (0) 3641 64 0 fax. 0049 (0) 3641 2856 but maybe this is the planetarium?
For the Oberkochen site: tel. 0049 (0) 7634 0 (this seems a little short?) fax. 0049 (0)7634 20 6808
Dear Fellow Microscopists: I would highly appreciate titles (and ISBN, address for ordering, possibly price..) of books and/or atlases on normal and pathological SEM material. I'd like to use them for teaching (first myself) and diagnostic purposes. Any suggestions would be welcome. Thanks in advance Peter P. Molnar molnarp-at-lib.dote.hu
Thanks for the responses. But it is apparent that I was not clear in stating my problem. So I will give it another try.
I am trying to dissect out some very small {50 um blood vessels. The main problem is that I cannot see them once the blood has washed out. I would like to stain the vessels with something that is compatible with immunohistochemical procedures using confocal microscopy. I am interested in immunostaining for the innervation and then determining, among other things, length density. Since most of the nerve are very small I would like to image them using the confocal microscopy. However, I am concerned that the vessel staining will interfere with the fluorescence when viewed on the confocal either by scattering or autofluorescence. Since alkaline phosphatase is present in most vessels and is still active after paraformaldehyde fixation, this should would work for the vessel stain. I am using rats and most likely the method of Mayahara et al., Histochemie 11:88;1967. I hope that this has clarified my problem and once again, any advice would be appreciated.
If you are fixing by perfusion,2% Evans blue in the fixative stains the vessels without any conflict with horseradish peroxidase or for that matter the fixation. I cannot comment on its fluorescence. Kate Connolly
To: Stephen J Murray {smurray-at-u.Arizona.EDU} Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com} *** Reply to note of 10/03/96 00:25
Stephen,
We have an S250 Mk II, also. I've got the image store unit on ours. It has a standard TV output (NTST). If you ask the people at Leo, they might be able to find you an old used one, for a decent price, considering how old it is. BTW, it's the HRRU, "high resolution record unit."
We have a thermal printer attached to the NTST output, but the image quality is poor. The HRRU has a resolution of 2500 verticle lines, and type 52 Polaroid film is about 1500 lines. The NTST should be about 350 lines. Thermal prints can be very inexpensive, but the resolution will never be very good.
At one time we had a data acquisition board attached, and took images from the visual display unit (don't remember line counts). The problem was software for handling the image capture and processing. A rudimentary version was developed locally, but then the support was dropped.
There are many commercial people in this business now. PC based image capture should be the best. I'm sure one can help you, but it will be costly. I'd like to know what the best solution is, since I'm not happy with ours, but our budgets are now very tight, too. Digitized images can be electronically distributed and processed as well. These are the reasons I have renewed interest.
Please keep this discussion public. I'm sure there are many people interested in this.
} If you are fixing by perfusion,2% Evans blue in the fixative stains the vessels } without any conflict with horseradish peroxidase or for that matter the } fixation. I cannot comment on its fluorescence. } Kate Connolly } Hello,
The Evans blue will emit very strong fluoresence in red but not green. We use it as a nice counterstain when using FITC as our primary tag.
Bob morphology core dermatology Univ. of Wash. Seattle
I would appreciate some pointers from anyone who is familiar with fixation of cultured cells to be subsequently used immunogold labeling. I have tried using 4% paraformaldehyde, 0.1% glutaraldehyde in PBS -at- pH7.4 but the cell morphology was quite chewed up. Thanks for your help.
Philomena Kaan UBC Pulmonary Research Laboratory St. Paul's Hospital Vancouver, B.C.
We are interested in making polyclonal antibodies to a 11 Kd purified protein. Do anyone have a recommendation of a company to use or to stay away from?
Patty Jansma Tel:602-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
Regarding your e-mail message of September 28/96 and your interest in the NanoLab 7 SEM:
Servicing, parts, spares and technical details are available from Semoptics Ltd. (613) 727-1698 (Voice or Fax). We are staffed by former employees who designed and manufactured the SEM at Semco Instruments in Ottawa, Canada from 1977-1984. The resolution of the NL7 with Tungsten emitter was {= 70 Angstroms.
Much more information by contacting Semoptics at the above phone number. Look forward to talking with you.
} Date: Thu, 3 Oct 1996 11:47:31 +100 } From: Dr. Molnar Peter {molnarp-at-lib.dote.hu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: SEM atlas } } Dear Fellow Microscopists: } I would highly appreciate titles (and ISBN, address for ordering, } possibly price..) of books and/or atlases on normal and pathological } SEM material. } I'd like to use them for teaching (first myself) and diagnostic } purposes. Any suggestions would be welcome. } Thanks in advance } Peter P. Molnar } molnarp-at-lib.dote.hu } Tissues and Organs: A Text-Atlas of Scanning Electron Microscopy 1979, R. G. Kessel and R H. Kardon W. H. Freeman and Co., San Francisco
ISBN 0-7167-0091-3 0-7167-0090-5 pbk
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
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At 07:51 2/10/96 -0400, you wrote: } } } On Tue, 1 Oct 1996, Probing & Structure wrote: } } } At 20:00 30/09/96 -0400, you wrote: } } } I have recently been using a Hitachi S-570 SEM. I wanted to use Polaroid 53, } } } but this being an older machine it does not have a setting for 800 speed } } } film. I was considering using the recommended setting for 400 ISO film and } } } close the lens aperture down one stop. Does anyone know if this is feasible } } } or can they recommend a better method. } } } } } } Thank you } } } Tom Bryner } } ****************************** } } Hi Tom and whoever: } } I believe that you cannot do better than to close down the camera } } by one stop. The only other consideration is lens resolution. For that size } } format best resolution for most lenses would be between f5.6 to 11. Lenses } } designed for 35mm format are likely to have best resolution with the } } aperture one stop more open. } } If the image is properly in focous, the additional depths of field } } is no consideration when photgraphing a flat screen. } } Cheers } } Jim Darley } } Probing & Structure } } } (Microscopy Supplies & Accessories) } } PO Box 111, Thuringowa QLD 4817 Australia } } } } } } Phone +61 77 740 370 Fax: +61 77 892 313 } } } A great microscopy site http://www.ultra.net.au/~pns/ } } } Years ago I did a study on the appropriate F stop to set the camera on the } image recording unit for the SEM (JEOL-35). That instrument had a F4.5 } lens and the ability to adjust the contrast and brightness independently. } } The end result was interesting. The quality parameter measured was the } number of line pairs resolved on the photographic image (Polaroid type 55 } was used since it has the resolution to see 2000 line pairs over the 4X5" } image. The best result was F8. At smaller F stopsl (F11 and lower) the } brigtness had to be increased which had the result of blooming- the } phosphor was being pushed too hard. At larger F stops there was a lack of } depth of field field resulting in the outer corners of the image being out } of focus. Also noticed was defocusing of the image due to "mottling" of } the negative material. } } Incidentally, our MRS-3 standard has patterns to help check CRT } resolution. } } } Hope this little tutorial helps. } } Joe Geller } Geller Microanalytical Lab } Topsfield, MA 01983 } www.gellermicro.com *************************************************
Joe Geller's observations, no doubt were right. But he had to stop down the large format lens to F8 to overcome spherical aberrations. Defocussed edges indicate that and not a lack of depth of focous. Joe (and many SEM'ists) have a crummy lens on their SEM. The better solution is a good lens; but SEM manufacturers save money with cheap lenses and would argue that the resolution is good enough, considering the film format and SEM screen resolution. The result is that users pay for roll film and obtain an image quality which is no better than is attainable with 35mm format and a good macro lens. 50 ASA 35mm film easily resolves all SEM screen details. The beauty of such 35mm system is that the camera can be motorised and integrated with the SEM. This totally avoids double and unexposed negatives. I've done and it works well. Jim Darley
Congrats on getting an SEM into a school environment!
I kept this email that appeared on the listserver a few months ago for just such an occasion. I hope it's helpful. I left the author's details at the bottom for you to direct further questions. I'd suggest that if you go this way (i.e. on a Mac) that you get hold of the public domain software 'Image' from NIH - it's a very handy Image Processing/Image Analysis package.
Another option might be the frame grabber we have on our Cambridge S120 called 'Image Slave' which slots into a PC, and I'd get hold of Image Tools, another pub.dom. IA/IP program for PC's. To find an 'Image Slave' distributor in the US you might ask Steve Wisbey of OED Pty Ltd here in Oz (sbwisbey-at-ozemail.com.au).
I realise that money it a key issue but I'd strongly recommend trying to get a Back-Scatterd Electron detector at some stage. Kids love looking at hairy biological things which are buggers for charging under the beam which produces crap Secondary Electron images. The image from a BSE detector will not be troubled by charging 99% of the time.
Good Luck,
Geoff
:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: Geoff Avern Manager Microscopy Laboratories Australian Museum Email: geoffa-at-amsg.austmus.oz.au 6 College St Ph: (61)(2) 9320 6198 Sydney, Australia. 2000 Fax: (61)(2) 9320 6059 ::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
------------------------------------------------------------------------ From Hasso Weiland:
} From the feedback I obtained to my previous posting it seems that there is astrong interest for home made systems for digital imaging on analog SEMs. Here is my cooking recipe for digital imaging on a JEOL840 using a MacIIci. The cost of the whole system was about US$ 700 and two days of work.
Ingredients:
1 MAC ADIOS IIJr from GW Instruments, 35 Medford St. Sommerville, MA 02143, Tel: (617) 625-4096
1 MAC ADIO ABO (Analog Breakout box)
1 9ft cable with a 14 pin connector on the microscope side, loose ends to connect to the terminal on the ADIOS board. The cable connects on the microscope side to connector JA2 on the rear panel of the JEOL840. This connection will control the SEM. A BNC cable connects to the JEOL connector NA8. This cable will carry the video signal to one of the AD channels of the MACADIOS board. The video signal on NA8 carries the same signal which will be displayed on the SEM screen, thus whatever signal is selected for the SEM display, BSE, SE, or ??, will be recorded by the digital imaging sytem.
1 small software, controlling the beam. It will deflect the beam and read in the Video signal at each position. The resolution can be set by the user.
Pin connections on the JA2 connector are as following:
Pin 1: X-deflection connected to DA out (CHN0) Pin 2: Y-deflection connected to DA out (CHN1) Pin 4: Blank (blanks the SEM screen) connected to one of the Digital Out channels Pin 5: Relay (controls the relay in the JEOL840 for external scan coil control, (this relay is build in each microscope), connected to one of the Digital Out channels Pin 7, 14: Ground connected to Common
The board should be set up to 10V on the DA channels. 0 Volt is in the screen center. For the X-deflection, +9 Volts is on the left of the screen (when sitting in front of the microscope), -9 Volt on the right side. For the Y-deflection, -9 Volt is the top of the screen, and +9 Volt is the bottom.
For external control, set the Digital Out which is connected to Pin 5 to high, low will give the control back to the JEOL system. Blank (connected to Pin 4) is on when set to high, setting this Digital Out to low will leave the beam on (be careful, this can easily burn a black mark on your screen, always watch your screen when setting up the system, best is to turn brightness and contrast down).
The small C-code we are using is part of an other application (BKD analysis), thus it is not stand alone and not necessarily helpful to everyone. If interested, I can e-mail the code later to those who would like to have it as a guide for their own development.
If anybody needs more help, please let me know.
Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069, USA 412 337-3133 weiland_h-at-atc.alcoa.com
Our company E.L.I. sprl (Belgium) has developped a vey powerful system for electronic microscope grabbing: ORION 4.1.
We are sure you will understand how easy it can be to upgrade your SEM by acquiring digital pictures with such a convivial package.
For your information, the PC minimum requirements to run ORION 4.1 are the following: * Pentium 100 Mhz - 16 Mb RAM * ISA slot 16 bits full length * VGA graphic card 1 Mb
The price for the ORION 4.1 package (hardware card + software) is 8.000-US$.
The "installation" includes: * the physical installation itself and the tests * a full training for the users * one year hot line guarantee (in case of on site service, the travelling costs will be charged)
The price for this service depends on the country (do we have a distributor or not) and/or the technical capability of the customer to install itself the connection with on line help (phone, fax or Email) from our technical service.
Don't hesitate to contact me for more information and/or have a look to our http site: http://www.microscopy-uk.org.uk
At 09:01 03/10/1996 EDT, you wrote:
} } There are many commercial people in this business now. PC based } image capture should be the best. I'm sure one can help you, but } it will be costly. I'd like to know what the best solution is, } since I'm not happy with ours, but our budgets are now very } tight, too. Digitized images can be electronically distributed and } processed as well. These are the reasons I have renewed interest. } } Please keep this discussion public. I'm sure there are many } people interested in this. } } . { } } { { } } } ===========} Dave King { { { } } } } ------------------------------------------------------ { { { { } }
Best regards,
Paul Vanderlinden. Sales Manager.
========================================================================= To contact us:
I suggest that you increase the concentration of glutaraldehyde to 1%. (4% PFA + 1% GA). When I fix cultured cells, I heat the fixative to 37 degr. C, and fix the cells taken directly out of the chamber -after one wash with buffer of the same temperature -(if they're not free-floating). I let them cool down in RT for a while, and then put them in the fridge for a couple of hours / over night. On the other hand; when you embed the cells in plastic that is suitable for immuno-gold staining + omit the osmium treatment, you don't really see what you're used to...
Good luck!
Margareta.Halin-at-ah.slu.se Dept. of Anatomy and Histology Sveriges LantbruksUniversitet
A post-doc or visiting scientist position is available at the University of Washington in Materials Science & Engineering for studies of metal - oxide interfaces using TEM and ELS. Other duties include help with supervision of TEM operation and general EM maintenance.
For further information please send a brief resume by email to -
Dr. R. M. Fisher Director - Electron Microscopy Consortium Univ. of Washington, Seattle, WA
Attached you will find a sketch of the Leeuwenhoek microscope scanned as a high resolution tiff image (slightly less than 500 kb when uncompressed). Legend to the figure:
a: Base b: Lens c: Height adjustment (coarse) d: Specimen holder and height adjustment (fine) e: Focusing screw
************************************************************************* Kristof KOVACS Associate Professor University of Veszprem, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684 *************************************************************************
We went over this long ago. NEVER, NEVER, post images compressed or otherwise to the listserver. If you wish to send an image to an individual that has requested it that is fine. But do not fill the worlds mailboxes with images, epecially using this forum. Too many people will have their mailboxes filled and it will result in bouncing Email messages (to me!) saying that there is a problem with user XYZ receiving Email.
If needed, I can and will supply FREE space on the MSA Anonymous FTP Site as well and my ANL Site in the Image Library sections for miscellaneous non-copyrighted and free/public domain images.
We went over this long ago. NEVER, NEVER, post images compressed or otherwise to the listserver. If you wish to send an image to an individual that has requested it that is fine. But do not fill the worlds mailboxes with images, epecially using this forum. Too many people will have their mailboxes filled and it will result in bouncing Email messages (to me!) saying that there is a problem with user XYZ receiving Email.
If needed, I can and will supply FREE space on the MSA Anonymous FTP Site as well and my ANL Site in the Image Library sections for miscellaneous non-copyrighted and free/public domain images.
Message-Id: {1.5.4.32.19961004124452.006a5154-at-biotech.ufl.edu} X-Sender: sdw-at-biotech.ufl.edu X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Nestor keeps an archive of everything on the list at the MSA home page "http://www.msa.microscopy.com/" Go to the "Microscopy and Microanalysis Software Library" Click on the "pub" directory and then on the "MicroscopyListserverArchives" directory. In ther you will find everything from about 1994 to current. It is not convienient nor searchable, but it is all there. Another option is "Tips & Tricks" at the web address listed at the end of this message. I maintain a biologic archive of the info posted to this list since about late 1994. It is more convient to use but limited to what I wanted to put in. If there is something you are specifically looking for or something you remember seeing recently but can't find, let me know. So far the archive is rather labor intensive and is usually about a month behind. I hope this helps.
At 05:08 PM 10/3/96 -0400, you wrote: } I believe I asked this before, if so, excuse this question: } } Is there a DIGEST mode for this list? } } Thanks. } Matt Stough } } } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
For cultured human colon adenocarcinoma cells, I use 2% freshly depolymerized paraformaldehyde (NOT formalin!) 70 mM NaCl 30 mM HEPES 5 mM CaCl2 pH 7.4
Cells like divalents (Ca2+) around during fixation. The osmolarity needs to account for the initial contribution of the aldehydes (a tricky situation since they are somewhat permeable but this increases with time. The infiltration into plastic should be gradual (2:1 and 1:1 solvent:resin).
} Hello All, } } I would appreciate some pointers from anyone who is familiar with } fixation of cultured cells to be subsequently used immunogold } labeling. I have tried using 4% paraformaldehyde, 0.1% } glutaraldehyde in PBS -at- pH7.4 but the cell morphology was quite } chewed up. Thanks for your help. } } Philomena Kaan } UBC Pulmonary Research Laboratory } St. Paul's Hospital } Vancouver, B.C.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
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Dear Confocal & Microscopy Listserv subscribers,
I'd like to make you aware of a suite of WWW pages that I've put together that touch on many of the topics related to biological microscopy and imaging. My target audience is students and staff (since that's who I work with the most), although I suspect "old hands" will find some useful links as well.
The main page is entitled "Microscopy and Imaging on the WWW". Related pages touch on Histology, Electron Microscopy, Confocal Microscopy, Digital Imaging and a page on free magazine subscriptions that might be of interest to microscopists.
The URL is (I had no control over how long this is): http://www.pharm.Arizona.edu/centers/tox_center/swehsc/exp_path/m-i_onw3.html
Your comments & feedback on this work in progress would be appreciated.
Yours,
Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: dcromey-at-ccit.arizona.edu) : :...................................................................: http://www.pharm.arizona.edu/exp_path.html
} Date: Fri, 4 Oct 1996 09:06:49 +0100 } From: Margareta Halin {Margareta.Halin-at-ah.slu.se} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: re: Fixation of cells for immuno-gold staining } } Hi! } } I suggest that you increase the concentration of glutaraldehyde to 1%. (4% } PFA + 1% GA). } When I fix cultured cells, I heat the fixative to 37 degr. C, and fix the } cells taken directly out of the chamber -after one wash with buffer of the } same temperature -(if they're not free-floating). I let them cool down in RT } for a while, and then put them in the fridge for a couple of hours / over } night. On the other hand; when you embed the cells in plastic that is } suitable for immuno-gold staining + omit the osmium treatment, you don't } really see what you're used to... } } Good luck! } } Margareta.Halin-at-ah.slu.se } Dept. of Anatomy and Histology } Sveriges LantbruksUniversitet }
However, be aware that glutaraldehyde can reduce or destroy some antigenic sites. The general rule is that the higher the glutaraldehyde concentration, the better the ultrastructure, but the worse the immunostaining. The only way to know if your particular antigen remains active after glut is to try it. Usual percents used range from about 0.1 to 1%, mixed with paraformaldehyde (4-8 %). Also, sometimes staining decreases with increased storage of specimens in aldehydes, even formaldehyde. If you're starting with an unknown, your best bet is to keep the glut low (we don't use it), and the fixation time short (1-2 hrs). Then you can test to see if more glut, longer fixation times, or even osmium fixation (not usually used except in freeze substitution), can be tolerated. Margareta's statement that the ultrastructure of non-osmicated tissue is not what you're used to seeing is very true. Maybe your cells aren't chewed up--just not stained conventionally.
Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
On Thu, 3 Oct 1996 PKaan-at-prl.pulmonary.ubc.ca wrote:
} Hello All, } } I would appreciate some pointers from anyone who is familiar with } fixation of cultured cells to be subsequently used immunogold } labeling. I have tried using 4% paraformaldehyde, 0.1% } glutaraldehyde in PBS -at- pH7.4 but the cell morphology was quite } chewed up. Thanks for your help. } } Philomena Kaan } UBC Pulmonary Research Laboratory } St. Paul's Hospital } Vancouver, B.C. } } Hello,
If the cells are to be post embed labelled I would worry about too strong of fixation in fear of loosing the immunolabelling capacity.
You might try a 2-4% para in sorensons (so there is no NaCl in the formula) or a combination of para and picric acid in sorensons buffer such as a Zamboni fixative, if you are post embed labelling on sections.
We have noticed a problem when there is NaCl present in the fix, with poor morphology.
On Thu, 3 Oct 1996 PKaan-at-prl.pulmonary.ubc.ca wrote:
} Hello All, } } I would appreciate some pointers from anyone who is familiar with } fixation of cultured cells to be subsequently used immunogold } labeling. I have tried using 4% paraformaldehyde, 0.1% } glutaraldehyde in PBS -at- pH7.4 but the cell morphology was quite } chewed up. Thanks for your help. } } Philomena Kaan } UBC Pulmonary Research Laboratory } St. Paul's Hospital } Vancouver, B.C. } }
You might try using a different buffer -- I've never had very good results fixing in PBS. We generally use 3% freshly (that's important) depolymerized formaldehyde and 0.05% glut. in a modified Hanks buffer. The more glut. your antigens will tolerate, the better. To get the membrane contrast closer to what you are used to seeing by conventional EM,try en bloc staining with Uranyl Acetate (which can help to improve the retention of antigenicity) and Tannic Acid (which can reduce antigenicity, so try some tissue +TA and some -TA). The composition of the modified Hanks fixative and protocols for en bloc staining can be found in:
J. Histochem. Cytochem. 40:845-857 '92.
There is (are?) a myriad of protocols for immunoEM. I sort of inherited this one and found it to work very well for many cell types and antigens, using either LRGold or Lowicryl K4M resins.
Happy Labeling!
Greg Martin gmartin-at-welchlink.welch.jhu.edu Dept. Cell Biology and Anatomy Johns Hopkins School of Medicine
Sara Miller wrote: } } On Fri, 4 Oct 1996, Margareta Halin wrote: } } } Date: Fri, 4 Oct 1996 09:06:49 +0100 } } From: Margareta Halin {Margareta.Halin-at-ah.slu.se} } } To: microscopy-at-Sparc5.Microscopy.Com } } Subject: re: Fixation of cells for immuno-gold staining } } } } Hi! } } } } I suggest that you increase the concentration of glutaraldehyde to 1%. (4% } } PFA + 1% GA). } } When I fix cultured cells, I heat the fixative to 37 degr. C, and fix the } } cells taken directly out of the chamber -after one wash with buffer of the } } same temperature -(if they're not free-floating). I let them cool down in RT } } for a while, and then put them in the fridge for a couple of hours / over } } night. On the other hand; when you embed the cells in plastic that is } } suitable for immuno-gold staining + omit the osmium treatment, you don't } } really see what you're used to... } } } } Good luck! } } } } Margareta.Halin-at-ah.slu.se } } Dept. of Anatomy and Histology } } Sveriges LantbruksUniversitet } } } } However, be aware that glutaraldehyde can reduce or destroy some } antigenic sites. The general rule is that the higher the glutaraldehyde } concentration, the better the ultrastructure, but the worse the } immunostaining. The only way to know if your particular antigen remains } active after glut is to try it. Usual percents used range from about 0.1 } to 1%, mixed with paraformaldehyde (4-8 %). Also, sometimes staining } decreases with increased storage of specimens in aldehydes, even } formaldehyde. If you're starting with an unknown, your best bet is to } keep the glut low (we don't use it), and the fixation time short (1-2 } hrs). Then you can test to see if more glut, longer fixation times, or even } osmium fixation (not usually used except in freeze substitution), can be } tolerated. Margareta's statement that the ultrastructure of } non-osmicated tissue is not what you're used to seeing is very true. } Maybe your cells aren't chewed up--just not stained conventionally. } } Sara } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735
Hello folks,
Here's a tip that will help your unosmicated/immunostained sections look more aesthetically pleasing. After you have immunostained your sections, presumably they are on nickel or gold grids, run the grids through a series of droplets which parallel a routine fixation: hard fix in 1-2% GA, buffer washes, OsO4, washes, UA, wash, blot dry and scope. You need only incubate for 10-15 min. in the fixes and 5 min. in the washes and so on. I've done this with LR White and Lowicryl and it works well. My colleagues see fine structure which more closely resembles "routinely" fixed TEM specimens and have their imunno-gold staining as well. Kind of like having your pie and eating it too. Try it - you'll like it!
Cheers,
John Aghajanian Worcester Foundation for Biomedical Research
Sorry for doing something wrong as . It was a coincidence that I saw the posting of Pete Barnett requesting an image of Leeuwenhoek microscope and just got a nice drawing of what is considered the very first useful microscope. Being happy with this finding I thought this clear drawing might be of interest to all microscopists considering the roots equally important to the newest findings or whatever. Sorry again, I will never do this in the future.
Kris
************************************************************************* Kristof KOVACS Associate Professor University of Veszprem, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684 *************************************************************************
I have been asked to give a presentation to a group of Microscope salespersons on PLM and specifically on the geological aspects. That is no problem, but some of their customers are ?biologists? or non-geological and are using some compensators in their research. They would like to know how to use them and what they are for. I can discuss how to make the appropriate measurements, but what materials are they measuring and what does that tell them? They are using Berek, Brace-Kohler, Erhinghaus and senarmont compensators. Anyone know what the ususal biological applications are? Thanks for any advice. You can e-mail me direct. If anyone else is curious, I can mail out a summary. Regards, Mike ================================================ Michael L. Boucher Sr. mboucher-at-isd.net 13345 Foliage Avenue Apple Valley, MN 55124-5603 Ph 612-432-8836 ================================================
Dear Stephen, There is a relatively inexpensive product to get an SEM image into a computer from GW Electronics, called Printerface. Larry Glassman of GW says it is a mail-order-type product, easy to install. Contact him at: GW Electronics 6981 Peachtree Industrial Blvd. Norcross GA 30092 Phone: 404-449-0707 Fax: 404-449-0284 I use a more expensive product, called PCI, and I find it is a wonderful way to show the SEM image to a class full of students or print out laser copies for all. Luck Mary.
Stephen Murray wrpte:
} I'm a high school teacher whose been given a vintage 1980 Cambridge } StereoScan 250 SEM. It is up and running as well as expected but now I } need to figure out how to get it more flexible. The recording device is a } Polariod affair that is much too expensive for numerous high schoolers. } What was intimated was hooking up a digitalizer to the TV mode of the } viewing screen and bypass the HRRT.(?) It was also told to me that if we } wanted to used the HRRT to capture the image digitally that it would take } a bit more money as well as smarts. (Two things I'm rather short on right } now). } } My mission (of which I am pleading for help with) is to find a product and } a vendor of such a digitalizer, a price, and all the pertinent information } to present at a meeting next week. } } Help please! } } Stephen Murray
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
The Second Edition of The Electron Microscopy Safety Handbook edited by Vernon C. Barber and Joseph A. Mascorro has been published for over a year. It is available from San Francisco Press, Inc., Box 426800, San Francisco, CA 94142-6800. I belive it is -at- $15-18. It has ISBN number of: 0-911302-72-7.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Thanks for all the suggestions on my question about adjustments necessary to the Hitachi S-570 when using an 800 speed film(Polaroid 53). The consensus of opinion was that changing the f stop is a viable method. Also it was suggested that calibration of the contrast and brightness would be necessary.
I changed the f stop from f16 to f22 with a scanning time of 100 seconds and adjusted the contrast and brightness settings. To compare I reduced the scan time to 50 seconds and used an f stop of f16. The resulting prints of these two tests were comparable with each other. I would say that they were slightly less sharp than using Polaroid 52 (ASA 400) at a scanning speed of 100 seconds and a f stop of f16, probably as to be expected. The results were acceptable though.
When time and circumstance allows a thorough calibration of the video signal will be done.
Many thanks for your program "EPCfocus". It is very good for teaching! May be you know some simulation programs for SEM? It is very interesting for me. I cannot link to programs "ChipSi" and "VBimage". Whatis the problem?
I look forward to hearing from you in the near future. Thanking you beforehand for you cooperation.
Whis best wishes, Dr. Igor Lapsker Center for Technological Education Holon Lapsker-at-barley.cteh.ac.il
Many thanks for your program "EPCfocus". It is very good for teaching! May be you know some simulation programs for SEM? It is very interesting for me. I cannot link to programs "ChipSi" and "VBimage". Whatis the problem?
I look forward to hearing from you in the near future. Thanking you beforehand for you cooperation.
Whis best wishes, Dr. Igor Lapsker Center for Technological Education Holon Lapsker-at-barley.cteh.ac.il
Thanks for answer my question refered to the tensile and torsion test by SEM, what you send me, was an important help to complete my thesis, thanks again
Is it possible to exclude subscribe and unsubscribe messages from the general messages that go out on the listserver? This would help keep down the traffic and save all of us from having to dump sll this extraneous mail!! Thanks, Steve
University of Wisconsin-Madison Robert M. Bock Laboratory 1525 Linden Dr rm#527 Madison, WI 53706
Work (608) 262-4581 Fax (608) 262-4570 Home (608) 837-9566
I have just finished equipping my lab with the essentials for TEM and finally I need to get the pictures out of the scope and onto print. Can anyone recommend a good professional enlargement system for 4 x 5 inch negatives?
Here is an update on the Cu concentration in the plant trichomes. Latest bulletin from trichome central is that the concentration is probably closer to 0.1% Cu.
Now that's a horse of a different color!
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
We need to digitize an entire 3.25 in. by 4.25 in. TEM negative and at the same time, maintain resolution adequate to distinguish the morphometry of the vesicles in synaptic terminals. Our CCD camera is unable to give us the resolution to distinguish the shape of the vesicles, so we were told that a flatbed scanner or a high resolution digital camera would solve our problem. We have an appointment for a demo of the digital camera but we also would like to have a scanner demo. Please let me know, if you are a representative who can demo a flatbed scanner for us or if you successfully use a flatbed scanner for resolution requirements similar to our needs. Thanks!
Donna Wagahoff SIU School of Med. P.O. Box 19230 Springfield, Il. 62794-1220
I agreed, particularly when the sender does not include a header or subject!
Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html Fax 504 587-7389 & Voice 584-2521, Internet: fermin-at-tmc.tulane.edu
Is it possible to exclude subscribe and unsubscribe messages from the general messages that go out on the listserver? This would help keep down the traffic and save all of us from having to dump sll this extraneous mail!! Thanks, Steve
University of Wisconsin-Madison Robert M. Bock Laboratory 1525 Linden Dr rm#527 Madison, WI 53706
Work (608) 262-4581 Fax (608) 262-4570 Home (608) 837-9566
I am trying to help a user of our lab solve a problem involving locating and quantifying small concentrations of copper in plant cells.
Here is the setup: Small plants are grown in soil high in copper. These plants accumulate copper in the trichomes (hairs) of the plant ( we think).
Here is the question: How can it be demonstrated that copper is actually accumulated in the trichomes and how closely can we approximate its concentration?
I have tried helping her by doing some quick and dirty SEM/EDS. I could not detect any copper in the trichomes (maybe I wasn't looking hard enough). She reports that there should be around 1% copper or less per dry weight of tissue. I suggested that that amount maybe hard to pin down using EDS, but I told her I would check it out with the experts to see what a reasonable minimum detectable amount might be (OK experts, what do you think?).
I also cautioned her regarding the pitfalls of EDS on biological samples, suggesting that she may want to explore alternatives to her preparation technique. She is currently fixing and embedding in paraffin so she can section for the light microscope. If freezing and other cryo techniques are the best way to go, I could use some help (and back-up) in advising her of how far to go and the chance of success. Is simple freezing enough or does she have to find someone with a cryo stage to keep everything in place throughout the analysis?
Finally, do you think there is any future in this type of investigation? If so, do you have any ideas of where we might be able to find the equipment needed to proceed? If not, do you have any alternatives you could share?
Perhaps I am too pessimistic, maybe I don't know what I am talking about. Either way it would help us to get some outside opinions and ideas about this problem.
Thanks,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Message-Id: {199610072241.RAA11685-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
If they are made to the proper server rather than to the "general" server, they won't go out over the listserver. Review the instructions that you received when you subscribed.
Joiner
At 11:09 AM 10/7/96 -0600, you wrote: } Is it possible to exclude subscribe and unsubscribe messages from the } general messages that go out on the listserver? This would help keep down } the traffic and save all of us from having to dump sll this extraneous } mail!! Thanks, Steve } } University of Wisconsin-Madison } Robert M. Bock Laboratory } 1525 Linden Dr } rm#527 } Madison, WI 53706 } } Work (608) 262-4581 } Fax (608) 262-4570 } Home (608) 837-9566 } } slimbach-at-facstaff.wisc.edu } } } }
Following Scott Walck's heads up on the TEM specimen prep. symposium, here is a repeat of the call for papers for the Materials Reliability in Microelectronics Symposium. Author instructions and meeting information are available at the web site listed at the end of this message. Abstract deadline is November 1, 1996.
Materials Research Society 1997 Spring Meeting March 31 - April 4, 1997 San Francisco, California
ABSTRACT DEADLINE: November 1, 1996
Symposium J: Materials Reliability in Microelectronics VII
The inexorable drive for increased integrated circuit functionality and performance places growing demands on the metal and dielectric thin films used in fabricating these circuits, as well as spurring demand for new materials applications and processes. In addition to meeting performance and manufacturability requirements, these materials and processes must yield circuits that operate reliably for many years. Achieving these goals requires a better understanding of the relationship of thin-film material properties and manufacturing processes to reliability degradation mechanisms.
This symposium is a continuation of the series that seeks to foster this understanding. The aim is to provide a forum to bring together researchers from industry and universities to discuss fundamental mechanisms and materials properties pertinent to materials reliability issues in the manufacture of submicron integrated circuits.
Papers are solicited in the following and related areas:
+ Reliability of metallic thin films and interconnects - Electromigration in lines, contacts, vias - Stress relaxation and stress voiding - Metal microstructure: grain growth, texture, precipitate formation - Effects of microstructure on electromigration - Intermetallic formation and diffusion barriers - New interconnect metallizations/novel alloys; alternative dielectrics + Reliability of dielectric thin films - Physics and chemistry of gate dielectric breakdown, including breakdown mechanisms, intrinsic vs. extrinsic failure, and failure analysis - Reliability, yield and gate stack processing: growth conditions, substrate effects, impurities, cleaning and plasma damage - Relation of reliability to leakage and tunneling currents, charge trapping and defect generation - Special reliability considerations for ultrathin ( { 5nm) gate dielectrics - Materials issues in hot-carrier reliability + Mechanical stress and strains in films, lines and device structures: deformation, fracture, adhesion, simulation and modeling. + Novel analytical and measurement techniques + Reliability modeling and simulations
Joint sessions are planned with Symposium I: "Polycrystalline Thin Films III: Structure, Texture, Properties, and Applications", and Symposium P: "Science and Technology of Semiconductor Surface Preparation"
A partial list of invited speakers: David Clarke (UCSB) W.W. Gerberich (University of Minnesota) Qing Ma (Intel) J. Meindl (Georgia Tech University) Wayne Paulson (Motorola) Klaus Schuegraf (Micron) M. Thouless (University of Michigan) A.H. Verbruggen (Delft University) W.L. Brown (Bell Laboratories) J.E. Greene (University of Illinois) C.V. Thompson (MIT)
Abstract information can be accessed through the MRS Website:
http://www.mrs.org/meetings/spring97/cfp/
Click on "abstract submission guidelines".
For further information, please contact Robert Keller (address, phone, email given below).
============================================================================== Robert R. Keller National Institute of Standards and Technology Materials Reliability Division, 853 office: (303)497-7651 325 Broadway FAX: (303)497-5030 Boulder, CO 80303 keller-at-boulder.nist.gov
Dear Jacob, The second edition was published a few years ago and should be available from the San Francisco Press. I think it is about $16. Jacob Bastack wrote: } Has the second edition of the Electron Microscopy Safety Handbook been } published? Luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Dear Jon, I think your only choice would be WDX, which has a lower detection limit than EDX. The sample, however, must be flat. A dressed block, such as is used to make ultramicrotome samples, may be suitable after carbon coating. Luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Message-Id: {199610080618.AA14010-at-lucy.swin.edu.au} Comments: Authenticated sender is {hbrinkies-at-gpo.swin.edu.au}
I have been approached by one of our PhD students to examine the surfaces of very small metal particles that are mounted in wax molds. However, I have never used the SEM for wax inpregnated specimens. Any advise is welcome. I only have two convential SEMs in my laboratory, and the preparation facilities are set up for metallographical and geological specimens, i.e. no cold stages and only a carbon evaporator.
Thanking you
Hans Brinkies Industrial Microscopy School of Mechanical and Manufacturing Engineering Swinburne, University of Technology Hawthorn - Australia
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} Greetings: } } I am trying to help a user of our lab solve a problem involving locating } and quantifying small concentrations of copper in plant cells. } } Here is the setup: Small plants are grown in soil high in copper. These } plants accumulate copper in the trichomes (hairs) of the plant ( we think). } } Here is the question: How can it be demonstrated that copper is actually } accumulated in the trichomes and how closely can we approximate its } concentration? } } I have tried helping her by doing some quick and dirty SEM/EDS. I could not } detect any copper in the trichomes (maybe I wasn't looking hard enough). } She reports that there should be around 1% copper or less per dry weight of } tissue. I suggested that that amount maybe hard to pin down using EDS, but } I told her I would check it out with the experts to see what a reasonable } minimum detectable amount might be (OK experts, what do you think?). } } I also cautioned her regarding the pitfalls of EDS on biological samples, } suggesting that she may want to explore alternatives to her preparation } technique. She is currently fixing and embedding in paraffin so she can } section for the light microscope. If freezing and other cryo techniques are } the best way to go, I could use some help (and back-up) in advising her of } how far to go and the chance of success. Is simple freezing enough or does } she have to find someone with a cryo stage to keep everything in place } throughout the analysis? } } Finally, do you think there is any future in this type of investigation? If } so, do you have any ideas of where we might be able to find the equipment } needed to proceed? If not, do you have any alternatives you could share? } } Perhaps I am too pessimistic, maybe I don't know what I am talking about. } Either way it would help us to get some outside opinions and ideas about } this problem. } } Thanks, } } Jonathan Krupp } Microscopy and Imaging Lab } University of California . . } Here is an update on the Cu concentration in the plant trichomes. Latest } bulletin from trichome central is that the concentration is probably closer } to 0.1% Cu.
} Now that's a horse of a different color!
Jonathon,
Even with the revised concentration estimate it MAY be possible to do by EM and possibly (?!) even without cryo.
Two key questions:
1. Is the Cu evenly distributed or concentrated in specific sights? If evenly distributed, then you have a difficult problem. If concentrated in very specific sights, you may have a somewhat easier problem. i.e. if it is 0.1wt% everywhere, you have a problem but the Cu may be concentrated in the surface layer of the hairs, then the LOCAL concentration could be much higher 1wt% - 5wt%.
So can you use knowledge of the metabolic routes involving the Cu to guess where it might be? Alternatively, BSE in the SEM or a STEM, or just high contrast in an unstained TEM section might indicate regions to check for Cu.
2. What makes you think that the Cu is still in the specimen? Unless you KNOW that the Cu has been immobilised, most specimen preparation procedures are not designed to fix elements but structure. So, unless I new different, I would assume that the Cu was washed out of the specimen during preparation. Although Cu does tend to be fairly immobile - it's not like Na or Ca.
IF the CU is immobilised AND locally concentrated, you stand a chance. but I would guess you still need to do something about the specimen preparation. Not being an expert in this area, I would suggest that you need to follow more standard EM preparation techniques but try to avoid all heavy metals (glutaradehyde fixation?) - their presence will mask small amounts of Cu. Of coure, this means that even if you do find the Cu, you may not be able to determine where it is in relation to the anatomy of the specimen!.
You also need to avoid coating the specimen - Au or C. In the SEM, this will probably mean low kV to minimise specimen charging but trying to pick up the Cu L peak will be difficult - high X-ray background at low energies - and you won't be able to go lower than about 2.5 keV anyway, or you'll never excite the Cu L peak. To excite the Cu K peak you'll need to be over 12keV and preferably around 20keV. All of this tends to suggest that you stand a better chance in a TEM.
More generally, if the Cu is mobile and/or not localised, to stand a chance of getting a result via EM, you will certainly need to follow a cryo route. This means freezing the specimen to fix the Cu in the correct place in the specimen and then using a cryo-stage to keep it cold.
My preference would be for cryo-TEM. The EDX spatial resolution will be higher - no beam spreading in a thin specimen - so IF the Cu is locally concentrated and you can find the right area, your chance of detection will be better.
A key factor in EDX of cryo-specimens is water content. If there is a lot of water in the specimen, this will obscure the presence of trace elements - you will need to dehydrate the specimen inside the EM. You'll probably need/want to do this anyway to improve resolution and contrast.
If cryo-SEM, don't coat the specimen - even with C and certainly not with Au or other heavy metals.
And don't forget that Cu is everywhere in most EMs. You need to do a careful check that any Cu you do pick up is really coming from the specimen - particularly in a TEM - e.g. Cu specimen grids, specimen holders (brass? and/or Cu parts), cold traps, an so on.
Also, I'd recommend a thorough literature search.
Hope that is some help - best of luck!
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: Larry-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
14TH INTERNATIONAL SPM WORKSHOP INTERNATIONAL WORKSHOP ON NEAR-FIELD SCANNING OPTICAL MICROSCOPY
Tuesday, October 22, 1996 8:00 a.m. to 6:00 p.m. Stanford University Stanford, California
International Workshop On Near-field Scanning Optical Microscopy Near-field Scanning Optical Microscopy (NSOM) allows the extension of optical microscopy, spectroscopy, and other photonic techniques beyond the Abbe barrier, also known as the diffraction limit. Recent advances in Scanning Probe Microscopy have made NSOM, first postulated in 1925, into a reality.
This workshop will showcase new applications and trends in NSOM and allow in-depth discussion with some of the world's leading practitioners of the technique.
NSOM HAS APPLICATIONS IN ALL AREAS OF SCIENCE
Based on classical optical contrast and providing unique information on electronic properties of matter, NSOM has potential applications in all areas of research. Speakers are drawn from groups interested in practical applications of NSOM to areas which are limited by conventional optical resolution. Areas of discussion will range from biology and clinical research to semiconductor technology and materials science. All invited participants are keenly interested in developing new areas of application for NSOM.
WHO SHOULD ATTEND
Because the workshop will aim to spark discussion of both exciting and novel NSOM applications, anyone with any interest in Scanning Probe Microscopy or anyone who would like to extend existing light-based studies to higher resolution should attend.
POSTER SESSION
Anyone with research in the field of NSOM or more general areas of Scanning Probe Microscopy is encouraged to submit a poster. Display facilities will be available during morning and afternoon breaks and during the lunch-time discussion period.
HOW TO REGISTER
Please complete the registration form (page 6) and return it to TopoMetrix Corporation in Santa Clara, California. This registration fee covers the full day*s attendance including eight speakers, lunch, and refreshments throughout the workshop.
AGENDA
8:00 - Registration 8:45 - Welcome address by Professor Ronald Hanson, Department Chairman, Department of Mechanical Engineering, Stanford University. 9:00 - Mark Utlaut, University of Portland - Introduction to Near-field Scanning
Optical Microscopy 9:30 - Philip Haydon, Iowa State University - Opportunities for Near-field Microscopy in Biology 10:10 - Break 10:40 - Wolfgang Heckl, University of Munich - Scanning Probe Microscopy of DNA from Molecules to Chromosomes 11:20 - Shyamsunder Erramilli, Boston University - Scanning Near-field Infrared Microscope (SNIM) Based on a Free Electron Laser 12:00 - Lunch/Poster Session 1:30 - Kenneth Goodson, Stanford University - Laser Thermometry and Processing in the Near-field 2:10 - Walter Duncan, Texas Instruments - Near-field Optical Spectroscopy of Electronic Materials and Structures 2:50 - Break 3:20 - Nigel Cave, Motorola Corp. - Near-field Optical Microscopy and Spectroscopy of Semiconductors 4:00 - Ludwig Balk, University of Wuppertal, Germany - Device Characterization and Failure Analysis by Means of Near-field Scanning Optical Microscopy 4:40 - Final Discussion
BIOGRAPHIES:
PROFESSOR LUDWIG BALK Dr. Balk studied physics at the Technical University of Aachen, gaining his diploma in 1971, then his doctoral degree within the faculty of electrical engineering in 1976. Following his move to Duisberg University as Academic Director in the Department of Materials, Dr. Balk took a full chair professorship for electronics in the faculty of electrical engineering at the University of Wuppertal in 1991 and is presently dean of faculty. His applications range from pure electronic materials to inorganic materials, such as functional ceramics, and biological materials, such as human tissues and cells.
DR. NIGEL CAVE Dr. Cave is a senior staff scientist in Material Research and Strategic Technologies at Motorola. He uses optical techniques to characterize new semiconductor materials
and processes. He is currently developing near-field optical techniques within Motorola to permit deep sub-micron analysis of semiconductor devices. Dr. Cave received his Ph.D. from Imperial College, London in 1990 and did his post-doctoral work at the University of Cincinnati.
DR. WALTER DUNCAN Dr. Duncan is a senior member of the technical staff in the Materials Science Laboratory of Texas Instruments. He received his B.S. in Chemistry from Virginia
Polytechnic Institute in 1974 and his Ph.D. in Inorganic Chemistry from North Carolina State University in 1979. His current research is directed towards in situ wafer state sensors for adaptive control of vacuum processes, near-field optical
microscopy, optical properties of quantum confined structures, and silicon based
optoelectronic integrated circuits.
DR. SHYAMSUNDER ERRAMILLI Dr. Erramilli is a professor in Photonics and Physics at Boston University, granted a joint appointment in 1996. He received his Ph.D in Physics from the University of Illinois working with Hans Frauenfelder. He later joined the faculty of the Physics department at Princeton University, where he taught for ten years. His research interests have included the application of imaging techniques in Biological Physics, using x-ray diffraction and high pressure to study biological systems. Most recently, Dr. Erramilli has worked in the development of new forms of infrared microscopy.
In 1995, he was named a recipient of the duPont Young Professor award.
PROFESSOR KENNETH GOODSON Dr. Goodson is a professor and Terman Fellow with the Mechanical Engineering Department at Stanford University. He received a Ph.D. from M.I.T. in 1993 and was visiting scientist for two years with Daimler-Benz AG. His research investigates thermal phenomena in semiconductor devices and interconnects for integrated circuits. Dr. Goodson's group is using NSOM to improve the spatial resolution of
far-field optical thermometry and thermal processing techniques developed previously in his laboratory. He is an Office of Naval Research Young Investigator and a recipient of the National Science Foundation CAREER Award.
PROFESSOR PHILIP HAYDON Dr. Haydon is a professor and the director of the Signal Transduction Training Group as well as the Laboratory of Cellular Signaling at Iowa State University. His research focuses on ion channels and synaptic trans-mission between neurons. He is applying AFM and NSOM techniques to these problems.
PROFESSOR WOLFGANG HECKL Dr. Heckl is a professor in Experimental Physics at the University of Munich in the Institute for Crystallography & Mineralogy. Dr. Heckl gained his Physics diploma at Munich in 1985 followed by his Ph.D. in experimental physics under Professors Moewald and Sackmann. Post-doctorate positions followed at the Max-Planck Institute and the University of Toronto before working with Professor G. Binnig at IBM Research (STM) and Prof. T. Hansch at the Ludwig-Maximilians University Munich (STM/SFM/NSOM).
PROFESSOR MARK UTLAUT Dr. Utlaut is a professor in the Department of Chemistry and Physics at the University of Portland where he pursues research in the applications of Focused Ion Beam and Secondary Ion Mass Spectrometry technology as well as NSOM imaging. Dr. Utlaut received his B.S. in Physics from the University of Colorado and his Ph.D. in Physics from the University of Chicago.
REGISTRATION HOW TO REGISTER Complete the advanced registration form below and return with your check or money order. Payment must be included in order to process your registration. On-site registration will be available on a first-come, first-serve basis only. All workshop registrations received by October 10, 1996 will be confirmed. Badges will be available at the workshop registration area on the day of the workshop. Registration begins at 8:00 a.m.
CANCELLATION AND REFUND POLICY If you need to cancel for any reason, you must notify TopoMetrix Corporation, in
writing, by 5:00 p.m., October 15, 1996 to receive a full refund. Because the tickets may sell out before the workshop, refunds will not be granted on cancellations received after that date.
SPEAKER AVAILABILITY The speakers listed in this brochure are leading professionals in their respective fields. Should a speaker be unable to attend the workshop, all efforts will be made to replace that speaker with one of comparable experience and qualifications.
WORKSHOP REGISTRATION: FAX to: 408/982-9751
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We invite and welcome anyone who is interested in visiting & seeing all of Polaroids equipment and supplies in Montreal, Quebec, Canada on October 9 & 10.
Anyone interested in a visit through our Polaroid Expo, please respond ASAP to polaroid-at-scottscientific.com
Scott Scientific _______________________________________________ SCOTT SCIENTIFIC P.O. Box 66552 Station Cavendish Montreal, Quebec, H4W 3J6, Canada
Assuming that the metal particles of interest actually protrude from the wax surface (and are not coated with wax), you should be able to image these samples and perhaps even do some EDS work on them if they are large enough (i.e. minimum of 1 micron). A thermal evaporated carbon film of about 10-15 nm should be sufficient for conductive purposes, however I would also paint a very thin trail of carbon dag from the surface of the sample along the edge to the SEM stud. You may have to image at lower kV's (10 or perhaps even 5 kV). If you find the sample still charges a little, try going down maybe 1kV and leave it in the same location for a few minutes. Sometimes you can balance the charge buildup and dissipation this way.
If your microscope has an auto brightness control, switch to manual control; and if you have digital imaging on the SEM, this will be a tremendous help as well in reducing the amount of charge observed in your final image, because images can be captured quickly.
Hope this is useful. If not, pretend I didn't write it.
Regards,
-Bob ****************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com ******************************* ______________________________ Reply Separator _________________________________
I have been approached by one of our PhD students to examine the surfaces of very small metal particles that are mounted in wax molds. However, I have never used the SEM for wax inpregnated specimens. Any advise is welcome. I only have two convential SEMs in my laboratory, and the preparation facilities are set up for metallographical and geological specimens, i.e. no cold stages and only a carbon evaporator.
Thanking you
Hans Brinkies Industrial Microscopy School of Mechanical and Manufacturing Engineering Swinburne, University of Technology Hawthorn - Australia
If I understand the requirements of this assay correctly, it seems to me that microscopy isn't necessarily the best technique to use in this application. I am not a biologist, but if you can somehow isolate/section away the trichomes from the plant tissues, you might be able to perform this analysis by AA (atomic absorption spectroscopy) or GFAA (graphite furnace AA techniques). Both of these methods yield detection limits far lower than you will ever get with EDS. I have performed similar analyses on tissues of marine bivalves with very good results.
Good luck in your studies;
-Bob ************************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com **************************************
I am trying to help a user of our lab solve a problem involving locating and quantifying small concentrations of copper in plant cells.
Here is the setup: Small plants are grown in soil high in copper. These plants accumulate copper in the trichomes (hairs) of the plant ( we think).
Here is the question: How can it be demonstrated that copper is actually accumulated in the trichomes and how closely can we approximate its concentration?
I have tried helping her by doing some quick and dirty SEM/EDS. I could not detect any copper in the trichomes (maybe I wasn't looking hard enough). She reports that there should be around 1% copper or less per dry weight of tissue. I suggested that that amount maybe hard to pin down using EDS, but I told her I would check it out with the experts to see what a reasonable minimum detectable amount might be (OK experts, what do you think?).
I also cautioned her regarding the pitfalls of EDS on biological samples, suggesting that she may want to explore alternatives to her preparation technique. She is currently fixing and embedding in paraffin so she can section for the light microscope. If freezing and other cryo techniques are the best way to go, I could use some help (and back-up) in advising her of how far to go and the chance of success. Is simple freezing enough or does she have to find someone with a cryo stage to keep everything in place throughout the analysis?
Finally, do you think there is any future in this type of investigation? If so, do you have any ideas of where we might be able to find the equipment needed to proceed? If not, do you have any alternatives you could share?
Perhaps I am too pessimistic, maybe I don't know what I am talking about. Either way it would help us to get some outside opinions and ideas about this problem.
Thanks,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
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---------- Forwarded message ----------
} Date: Wed, 2 Oct 1996 15:00:39 -0700 (PDT) } From: Sarah Bottjer {bottjer-at-alnitak.usc.edu} } To: neuroethologists {neuroethology-at-sio.ucsd.edu} } Cc: soumya iyengar {iyengar-at-usc.edu} } Subject: agarose embedding for immuno staining } Mime-Version: 1.0 } Status: RO } } does anyone out there have a good protocol for embedding tissue in agarose } (for subsequent frozen sectioning and free-floating immuno reaction)?? We } prefer not to use gelatin-albumin, since we are embedding small bryozoan } larvae, and it's a distinct advantage to be able to see them in the } agarose! } (p.s. please don't ask me how a songbird lab got involved with } bryozoans . . .) } Thanks in advance, Sarah Bottjer }
It is said, that the FT of drift is different to the FT of vibration (for example on carbon film only).
The reason for this difference is not clear to me, because in both cases the "points" on the object are represented on the image by an elongated shape.
Where does the difference in FT come from, and how does the difference in FT look like??
------------------------------------------------------------------------ Bettina Fedtke fedtke-at-ubaclu.unibas.ch Maurice E. Mueller Institute University of Basel, Switzerland Klingelberstr. 70 Tel +41-61-267 2257 CH-4056 Basel Fax +41-61-267 2259 ------------------------------------------------------------------------
At 07:08 PM 10/4/96 BST-1, Brian Ford wrote: } } } We have a new web page. You will find it on: } } www.sciences/demon.co.uk } } We have e-mail on demon, too: } } brian-at-sciences.demon.co.uk } } Let me know what you think of the web page - it's only provisional, I } shall not really announce it 'til Christmas. } } Best wishes, } Brian J Ford } Dear Brian,
If www.sciences/demon.co.uk is the correct address, than I cannot find it; I have tried on a number of occasions.
I do not think agarose freezes very well. The water tends to separate from the colloid and makes large ice crystals. At least this is what happens during slow freezing. On thawing the agrose then sorta precipitates out leaving free water. Way back when, we used to recycle agarose for microbiological culture and that is how we separated the agarose from the soluble components. I am trying to think of an insoluble alternative that might work. Maybe polyacrylamide } } ---------- Forwarded message ---------- } } } Date: Wed, 2 Oct 1996 15:00:39 -0700 (PDT) } } From: Sarah Bottjer {bottjer-at-alnitak.usc.edu} } } To: neuroethologists {neuroethology-at-sio.ucsd.edu} } } Cc: soumya iyengar {iyengar-at-usc.edu} } } Subject: agarose embedding for immuno staining } } Mime-Version: 1.0 } } Status: RO } } } } does anyone out there have a good protocol for embedding tissue in agarose } } (for subsequent frozen sectioning and free-floating immuno reaction)?? We } } prefer not to use gelatin-albumin, since we are embedding small bryozoan } } larvae, and it's a distinct advantage to be able to see them in the } } agarose! } } (p.s. please don't ask me how a songbird lab got involved with } } bryozoans . . .) } } Thanks in advance, Sarah Bottjer } } } } } } ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
Both drift and vibration will smear out image points into line segments. The image power spectrum will then be truncated in the direction of the drift or vibration. How can we distinguish between drift and vibration in the power spectrum?
Theoretical answer: For an exposure time that is longer than several vibration periods, vibration smear will have a Gaussian-like profile. Drift smear will be constant over the exposure with an abrupt step at each end of the line segment. The FTs of these functions will be different; one will produce a reciprocal-space damping function with a Gaussian-like profile, whereas the other will have a sin(x)/x profile showing a banded ringing effect.
Practical answer: A series of images taken with increasing exposure times will show the same truncation of the power spectrum (FT) for vibration, but an increasing effect for drift (assuming the drift is fairly constant).
Mike O'Keefe -------------------------------------- drift and vibration
It is said, that the FT of drift is different to the FT of vibration (for example on carbon film only).
The reason for this difference is not clear to me, because in both cases the "points" on the object are represented on the image by an elongated shape.
Where does the difference in FT come from, and how does the difference in FT look like??
------------------------------------------------------------------------ Bettina Fedtke fedtke-at-ubaclu.unibas.ch Maurice E. Mueller Institute University of Basel, Switzerland Klingelberstr. 70 Tel +41-61-267 2257 CH-4056 Basel Fax +41-61-267 2259 ------------------------------------------------------------------------ :.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.: Michael A. O'Keefe, Deputy Head National Center for Electron Microscopy Lawrence Berkeley National Laboratory University of California Berkeley, California 94720 tel: (510) 486-4610 fax: (510) 486-5888 email: maok-at-lbl.gov http://ncem.lbl.gov/ :.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:
8:00-8:30 Registration, Vendor Displays and Welcome Coffee at Jesse Wrench Auditorium.
8:30-8:35 Welcome and comments.
8:35-8:55 Don Parker, Monsanto, St. Louis. "Network Interfacing of Microscopes"
8:55-9:15 Lee Dickey, Micro Lines Marketing, St. Louis. "Ultramicrotomy in Material Sciences"
9:15-9:35 Nancy Wilson Rizzo, SemTech, Inc, Shawnee Mission, Kansas. "Analysis of Gunshot Residue"
9:35-10:05 Phil Fraundorf, Physics and Astronomy, University of Missouri-St. Louis. "Some Developments in Air-Based Scanning Tunneling Microscopy"
10:05-10:30 Morning Break-Refreshments.
10:30-11:30 Dale E. Newbury (MAS Tour Speaker) National Institute of Standards and Technology, Gaithersburg, Maryland. "Lies, Damned Lies, and Standardless Analysis"
R.S.V.P. -- Lou Ross, (573) 882-4777 or geosclmr-at-showme.missouri.edu or Fax (573) 882-5458 by Tuesday, October 15th!
1:00-1:20 Heidi Schatten, Veterinary Sciences, University of Missouri-Columbia. "Cytoskeletal Organization during Fertilization, Cell Division, and Development in Microgravity: Results from a Space Flight (STS-77) on the Shuttle Endeavor"
1:20-1:40 Tobias Baskin, Biological Sciences, University of Missouri-Columbia. "Cryofixation of Large Samples for Light Microscopy"
1:40-2:00 Hari B. Krishnan, Plant Pathology, University of Missouri-Columbia. "Immunocytochemical Localization of Storage Proteins in Developing Rice Endosperm"
2:00-2:20 John Burton, Cardiology, University of Missouri-Columbia. "Utilization of Confocal Laser Microscopy and Image Analysis as Tools in Medicine Today"
2:20-2:35 Afternoon Break-Refreshments
2:35-2:55 James W. Cogswell, L.M. Ross Jr., M.J. O'Brien, H. Neff, and M.D. Glascock, Anthropology and Geological Sciences, University of Missouri-Columbia. "Postmanufacture Effects on the Chemical Characterization of Prehistoric Pottery: Evidence from the Central Mississippi River Valley"
2:55-3:15 Robert R. Church, Anthropology, University of Missouri-Columbia. "The Effect of Boiling on Osteological Materials"
3:15-3:35 Rajat Roychoudhury, E.J. Charleson, and L.M. Ross, Jr, Electrical Engineering and Geological Sciences, University of Missouri-Columbia. "Microcharacterization of Phosphorus Doped Diamond Thin Film Interfaces Using Voltage Contrast and Electron Beam Induced Current Studies"
3:35-3:55 Fariborz Golshani, M. Prelas, L.M. Ross, Jr., Nuclear Engineering and Geological Sciences, University of Missouri-Columbia and John L. Fitz, University of Maryland. "CL Study of Polycrystalline Diamond Films Before and After Annealing"
4:00- Business Meetings
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I have never worked with poly-L-lysine coated slips (but soon will) and have a couple of questions. It looks like a fairly easy procedure but none of the several dozen EM books on my shelf say much about it.
1. What is the shelf life of an aqueous solution of poly-L-lysine? 2. What is the shelf life of coated cover slips and how should they be stored? 3. When making lysine coated cover slips, the procedure (as I understand it) is to cover a cover slip with an aqueous solution of poly-L for one hour then dab dry. Can that drop of solution be transferred to a fresh cover slip or has its supply of lysine been exhausted? Do slips need to be cleaned in any particular fashion? 4. When working with cells in culture, is it better to stick them to the cover slips as a living culture or after glut or osmium treatment? 5. If living cells are exposed to a coated cover slip, does subsequent glutaraldehyde treatment cross link the cells to the lysine and increase adhesion? 6. How long should the cell culture be left on the coated cover slip? Temperature? 7. What density of cells in the culture works best? 8. Any special concerns during dehydration, CPD, or sputter coating? 9. Did I leave anything out?
Thanks in advance Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
} It is said, that the FT of drift is different to the FT of } vibration (for example on carbon film only). } } The reason for this difference is not clear to me, because in } both cases the "points" on the object are represented on the } image by an elongated shape. } } Where does the difference in FT come from, and how does the } difference in FT look like?? } Dear Bettina, Assume for the moment that the vibration has constant amplitude, the drift is always in the same direction and the exposure time for the image is long compared to the period of the vibration. In this case, the image corresponding to a point is a line segment which is of fixed length in the case of a vibration and which is proportional to the exposure time for drift. Furthermore, if the vibration is from simple harmonic motion, the intensity along the line segment is greater at the ends than in the middle; whereas, the intensity for drift is constant for constant drift velocity. I won't go into the details of the math, but there will be a large Fourier amplitude for the spatial frequency corresponding to the amplitude of the vibration or for the distance of the drift. There will be differences in the higher-frequency Fourier amplitudes depending on whether the intensity along the line segment is variable (vibration) or constant (drift), and for a series of different exposure times, these large amplitudes will occur at the same (vibration) or different (drift) frequencies. That said, in the real world, vibrations do not have constant amplitude, drift is not unidirectional and, depending on the source, exposure times are not necessarily longer than vibration periods, al- though they usually are. Another consideration for distinguishing drift from vibration by taking a series of exposures of different lengths is that the drift may be induced by the beam and may vary with the beam conditions. By spreading the beam to get the proper illumination for a longer exposure, you might slow the drift, so that the line segment produced will be the same length on the film. The easiest way to see what the differences in the FT look like is to rotate the image so that the axis of the line segments is along the x- or y-axis and do a 1-D FT; i.e. multiply the intensity by cos(2pi hx/l) and integrate over x, where l is the length of the image. This will give F(h) with the peaks as described above. The line segments should be uncorrelated, so the Fourier amplitudes from each should add with random phase, and the total amplitude distribution should look like that from a single line segment. Yours, Bill Tivol
2:00 - 2:45 Imaging Applications in the Pharmaceutical Industry: Not Just Pretty Picture {/fontfamily} {fontfamily} {param} Times {/param} s {/fontfamily} {fontfamil= y} {param} Times_New_Roman {/param}
Michail Esterman, Technology Core Research, Lilly Research Laboratories, Indianapolis I {/fontfamily} {fontfamily} {param} Times {/param} N
3:15 - 4:00 Electron Microscopy in Pathology: Under the Regulatory Microscop {/fontfamily} {fontfamily} {param} Times {/param} e {/fontfamily} {fontfam= ily} {param} Times_New_Roman {/param}
Sandy L. White and Jeffrey W. Horn, Toxicology Research Laboratories, Lilly Research Laboratories, Greenfield I {/fontfamily} {fontfamily} {param} Times {/param} N
4:00 - 4:45 Microscopy in Medical Product Developmen {/fontfamily} {fontfamily} {param} Times {/param} t {/fontfamily} {fontfa= mily} {param} Times_New_Roman {/param}
Jim DiOrio, Medical Materials Technology Center, Baxter Healthcare Corporation, Round Lake I {/fontfamily} {fontfamily} {param} Times {/param} L
The CHICAGO and CHICAGO WESTERN CHAPTERS Of ASM INTERNATIONAL
The MIDWEST MICROSCOPY andMICROANALYSIS SOClETY and The MATERIALS SCIENCE DIVISION of ARGONNE NATIONAL LABORATORY
presents
"Scanning Electron Microscopy Today" An All-Day Educational Seminar
Friday, October 25, 1996 Advanced Photon Source Conference Center Auditorium Argonne National Laboratory
8:30 am-4:30 pm
The lecturers for this seminar will review in some depth the basic principles of SEM, including the basic concepts of SEM operation and specimen preparation, imaging mechanisms in various operational modes, field emission SEM (FESEM), low voltage SEM (LVSEM) and environmental SEM (ESEM). The lecturers include Professor David Joy, University of Tennessee Knoxville (sponsored by Nissei Sangyo America/ Hitachi Scientific Instruments) and Vernon Robertson, JEOL Applications Laboratory (sponsored by JEOL USA, Inc.). Whether you are a beginner or a long-time user of scanning electron microscopy, you will find the lecturers and their lecture material interesting, informative and uptodate. Extensive discussion is encouraged. Registration fee is $20 per person (students, $10), which includes a buffet lunch and refreshments during morning and afternoon breaks and should be paid in advance. To register, complete and mail the form below, along with a check or money order for registration. If you are not a US citizen (even if you hold a Green Card), you must provide city and date of birth in order for us to arrange for your entry to the Laboratory site. Please register early and save everyone considerable hassle. The day of the Seminar, come directly to North Gate where the security guard will check your name on a list of attendees. Bring photo ID (e.g., drivers license).
At 07:08 PM 10/4/96 BST-1, Brian Ford wrote: } } } We have a new web page. You will find it on: } } www.sciences/demon.co.uk } } We have e-mail on demon, too: } } brian-at-sciences.demon.co.uk } } Let me know what you think of the web page - it's only provisional, I } shall not really announce it 'til Christmas. } } Best wishes, } Brian J Ford }
Maybe I'm wrong, but I consider the announcement of a webpage on such a large mailing list as an official announcement. Moreover, if it concerns a PROVISIONAL webpage (don't we all have one) I think it is something like announcing the submission of a paper.
Possibly it's better to announce these type of things on a smaller platform, like colleagues and friends.
If no one agrees with me I'm sorry to have bothered you.
We are undergoing renovation of our shared microscopy facility and need specific heat load information for each piece of equipment in order to convince the engineers of the need for dedicated air cooling/ventilation. Does anybody know the following or a reference for the following: The heat load for: 100W mercury lamp 100W halogen lamp Nikon incubator boxes NP-2 Olympus incubator boxes Sony 17" monitors (computer) Sony 14" monitors computers VCRs anything else that we may have overlooked (each person is calculated as 75W)
Mr-Received: by mta RANDD; Relayed; Thu, 10 Oct 1996 09:31:43 -0500 Mr-Received: by mta MCM$RAND; Relayed; Thu, 10 Oct 1996 09:31:44 -0500 Mr-Received: by mta RANDD; Relayed; Thu, 10 Oct 1996 09:32:08 -0500 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
Midwest Microscopy and Microanalysis Society Meeting
Microscopy: A Problem - Solving Tool in the Healthcare Industry
Tuesday, October 29, 1996 Abbott Laboratories Abbott Park, Illinois
12:00 - 1:00 Lunch (provided by Abbott Laboratories)
1:00 - 1:15 Welcome
1:15 - 2:00 Pharmaceutical Sleuthing - Applications of Light Microscopy Scott Aldrich, Trace Substance Analysis, Pharmacia & Upjohn, Inc.; Kalamazoo MI
2:00 - 2:45 Imaging Applications in the Pharmaceutical Industry: Not Just Pretty Pictures Michail Esterman, Technology Core Research, Lilly Research Laboratories, Indianapolis IN
2:45- 3:15 Break
3:15 - 4:00 Electron Microscopy in Pathology: Under the Regulatory Microscope Sandy L. White and Jeffrey W. Horn, Toxicology Research Laboratories, Lilly Research Laboratories, Greenfield IN
4:00 - 4:45 Microscopy in Medical Product Development Jim DiOrio, Medical Materials Technology Center, Baxter Healthcare Corporation, Round Lake IL
4:45 Tour of the Department of Microscopy and Microanalysis
To register, please contact Jerry Gagne by October 11 phone (847-938- 5023), FAX (847-938-5027) or E-mail (gerard.d.gagne-at-abbott.com)
Please help us out by registering ahead of time! The main security gate at Abbott requires a list of registrants in advance, and we need to plan for lunch. See you on October 29th!
Additional information will be posted on the MMMS WWW Site: http://www.msa.microscopy.com/MSALAS/MMMS/MMMSHomePage.html
} 1. What is the shelf life of an aqueous solution of poly-L-lysine? I would think only a few days. I made a concentrated solution up under sterile conditions, aliquoted it and froze it at -20. It lasts for 6 months if frozen. } 3. When making lysine coated cover slips, the procedure (as I understand } it) is to cover a cover slip with an aqueous solution of poly-L for one } hour then dab dry. Iwas using the coverslips for culturing work. I would coat the coverslips for 10 minutes, remove the PLL, rinse the coverslips 2X with deionized water and then let them dry. } Can that drop of solution be transferred to a fresh } cover slip or has its supply of lysine been exhausted? Partially exhasuted and you may want the same concentration of PLL to be on each coverslip.
Please let me take a moment to thank everyone who offered suggestions on how to solve our problem of localizing Cu in trichomes. We got some great ideas and will try to move forward on the project.
Also, let me express my gratitude to Nestor and each of you who read and contribute to this list. For me, this list represents a tremendous source of knowledge and help when I need advice. It provides a helpful reality check whenever I wonder if what I want to do is not quite thought out or needs some sage advice from those with more experience than I.
Thanks again,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Please let me take a moment to thank everyone who offered suggestions on how to solve our problem of localizing Cu in trichomes. We got some great ideas and will try to move forward on the project.
Also, let me express my gratitude to Nestor and each of you who read and contribute to this list. For me, this list represents a tremendous source of knowledge and help when I need advice. It provides a helpful reality check whenever I wonder if what I want to do is not quite thought out or needs some sage advice from those with more experience than I.
Thanks again,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
} We are undergoing renovation of our shared microscopy facility and need } specific heat load information for each piece of equipment in order to } convince the engineers of the need for dedicated air cooling/ventilation. } Does anybody know the following or a reference for the following: } The heat load for: } 100W mercury lamp } 100W halogen lamp } Nikon incubator boxes NP-2 } Olympus incubator boxes } Sony 17" monitors (computer) } Sony 14" monitors } computers } VCRs } anything else that we may have overlooked (each person is calculated as 75W) } } Thanks- } Michael Cammer You dont' go far wrong (and err on the safe side anyway) if you just take the wattage of the equipment as representing the heat load. In Australia we express heat load in watts anyway. If its different in the US a competent Air-conditioning engineer can convert for you.
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This waas sent to me byu a colleague and hopefully someone can help her. I have little experience doing double labeling. Thanks.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 //// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
---------- Forwarded message ---------- prutle1-at-umbc.edu, Tom Roth {roth-at-umbc.edu}
I have been trying to use the confocal microscope to determine when the chicken protein LEP100 arrives in the lysosomes of transiently expressing Drosophila tissue culture cells. I have not had much success, and would greatly appreciate any help or advice you could give me.
} From the experiment I usually see about 1-5% of the cells expressing LEP100. The amount of protein expressed can vary from cell to cell, depending on the copy number. To determine whether or not LEP100 reaches the lysosomes, I prelabel the lysosomes with Fluorescence Dextran. I then localize LEP100 by indirect immunofluorescence using RITC-conjugated secondary antibody. I see lysosomal labeling in the green channel and LEP100 staining in the red channel, but there intensities are different. There appears to be colocalization using the naked eye. THe problem comes in when I try to sequentially scan each image and superimpose the images. I don't see colocalization. That is I don't see yellow areas where the red and green areas overlap.
The procedure that I am using on the confocal microscope (LEICA TCS 4D) is as follows: I first select optical fiters for channel 1 (FITC, with excitation of 488nm). Next, I adjust the voltage, offset, and pinhole for optimal image quality in channel 1. This typically results in a voltage of 550, an offset of -2, and a pinhole of 100 (for the 100X objective). The laser power is about 9. I next define the top and bottom of the Z series, set the memory bar to the number of sections to be taken, and then scan the image doing an average of 4 scans per frame. Then I go back and select optical filters for channel 2 (RITC, with excitation of 568nm). I then adjust the voltage and offset for optimal image quality when in channel 2. This typically results in a voltage of 920 and an offset of -1. I do not change the dimensions for the Z series, laser power, or pinhole size. After scanning the same image series in channel 2, I go to the TCD monitor to display the images. TO get the best resolution, I just look at one section of the image series at a time. I go to the Macrohandling window and click on gallery all for 2 channels. What I see on the TCD display monitor is the image of one section in channel 1 (lysosomes), and the image of the same section scanned in channel 2 (LEP100) superimposed on top of the channel 1 image. I see green areas and red areas which appear to overlap, but the color is not yellow. Do you know what is going on? Please help me if you can. Thank you.
This is a request from L. H. Kuo. I am a electron microscopist. I would appreciate very much if you can transfer the information related to electron microscpy from your internet connection.
Thank you very much.
Best regards,
Li-hsin Kuo.
******************************************************* Joint Research Center for Atom Technology (JRCAT), c/o National Institute for Advanced Interdisciplinary Research (NAIR), 1-1-4 Higashi, Tsukuba, Ibaraki 305, Japan. Tel: 81-298-54-2739 Fax: 81-298-54-2777 *******************************************************
} Date: Thu, 10 Oct 1996 10:00:35 +0100 } From: Adriaan Houtsmuller {houtsmuller-at-pa1.fgg.eur.nl} } To: Microscopy-at-aaem.amc.anl.gov } Subject: Re: www } } At 07:08 PM 10/4/96 BST-1, Brian Ford wrote: } } } } } } We have a new web page. You will find it on: } } } } www.sciences/demon.co.uk } } } } We have e-mail on demon, too: } } } } brian-at-sciences.demon.co.uk } } } } Let me know what you think of the web page - it's only provisional, I } } shall not really announce it 'til Christmas. } } } } Best wishes, } } Brian J Ford } } } } Maybe I'm wrong, but I consider the announcement of a webpage on such a } large mailing list as an official announcement. Moreover, if it concerns a } PROVISIONAL webpage (don't we all have one) I think it is something like } announcing the submission of a paper. } } Possibly it's better to announce these type of things on a smaller } platform, like colleagues and friends. } } If no one agrees with me I'm sorry to have bothered you. } } A.B. Houtsmuller } } __ __ } __ /_/ /_/ } /_/ } ____ ___ } / / / | } / / / / *Dept. of Pathology, } / --- / *Erasmus University, } / / *P.O. BOX 1738, } /__ ** | *3000 DR Rotterdam, } / | *The Netherlands, } __/ | *Phone +31 10 4087499 } \________ / *Fax +31 10 4366660 } | \ } / | } ---- }
I agree. At least you could tell us in the subject heading what it's about. I, for one, don't have time to look it up just to see if I'm interested.}
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} We have a new web page. You will find it on: } http://www.sciences.demon.co.uk [corrected] } Brian J Ford
} Maybe I'm wrong, but I consider the announcement of a webpage on such a } large mailing list as an official announcement.... } A.B. Houtsmuller
I suspect that sending the message to the microscopy list was unintended, considering the long list of addresses on the original note. I would have ignored this except that I took the time to find out that the '/' should be a '.'. My apologies to those who can't rapidly scan over junk mail, but there are better mail readers available...
Anyway, readers of the list should be careful about putting the list address into mail aliases to which they might later send something that doesn't belong.
Just a short note to let you all know that I will be out of the country from Oct 12-20. This guarenttee's, of course, that SOMEONE SOMEWHERE will attempt SOMETHING that will upset the Listserver and send it into chaos (why should this trip be different than any others, right?). Unfortunately, my connection to the Net will not be very good so be prepared for a possible hickup that may not be immediately noticed by yours truely.
If your not sure of something today is the day to ask the question as I'll be leaving tomorrow, Saturday Oct. 12.
--------------
Just as a reminder to UNSUBSCRIBE
send a message to: LISTSERVER-at-MSA.MICROSCOPY.COM
the message type UNSUBSCRIBE MICROSCOPY "USERID-at-YOUREMAILADDRESS".
If you send these messages to MICROSCOPY instead of LISTSERVER, while I am gone I can almost guarentee you will NOT be unsubscribed since I will not be monitoring the postings and will NOT catch your errors.
If you think you will be unsubscribing during the next week, why not do it today to save possible hickups to the system while I'm gone.
---------------
Nestor Your Friendly Neighborhood SysOp
For those of you that are curious I will be at the Microscopy & Materials Conference (MicroMat - 96) in Rio de Janerio.
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Someone asked for estimated heat loads from laboratory equipment. An easy and reasonable approximation is to assume all power consumed is converted to heat. So that computer (w/200 watt power supply) and monitor (200 watts) will dump up to 400 watts into a room; don't adjust this estmate to reflect that portion of time the system is off or asleep because you want the HVAC to be able to handle the maximum loads.
1. Clean coverslips (first rinse with methanol, then water) 2. 1ug/ml polylysine solution (high molecular weight) 3. fixed cells 5. standard aldehyde solution
Spread a drop of the polylysine solution on a clean coverslip. Spread the drop by drawing the edge of one coverslip over the surface of the other. Allow the coverslip to dry for 1 min. Rinse with double distilled water. Place a drop of the sample that is in water or buffer on the reactive surface. Wait for the sample to settle down on the surface, usually between 5-20 min. Place coverslips in aldehyde solution for 15 min. Proceed as normal for dehydration and drying.
Coverslips must be clean or polylysine won't stick. Unfixed cells will adhere but they tend to flatten out on the surface. A light fixation before putting them on the coverslips helps prevent this occurrence. The polylysine I use has been in the refrigerator for several months. The container I used previously lasted almost two years. You want a good growth of cells. I make up my coverslips fresh.
Ginger R. Baker EM Lab Dept. of Anatomy, Pathology, and Pharmacology 250 Vet Med OSU Stillwater, OK 74078 (405) 744-6765 FAX: (405) 744-5275 Email: lizard-at-okway.okstate.edu
The header of a mail message being sent to you was damaged during transit, so could not be delivered normally. The full message is included below.
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Received: from Sparc5.Microscopy.Com by bham.ac.uk with SMTP (PP); Fri, 4 Oct 1996 06:17:05 +0100 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id VAA27821 for dist-Microscopy; Thu, 3 Oct 1996 21:34:48 -0500 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id VAA27818 for {microscopy-at-sparc5.microscopy.com} ; Thu, 3 Oct 1996 21:34:46 -0500 Received: from smtp.unsw.edu.au ([149.171.168.151]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id MAA05464 for {microscopy-at-sparc5.microscopy.com} ; Fri, 4 Oct 1996 12:39:00 +1000 (EST)
Good books with SEM pictures
Scanning Nature D. Claugher British Museum ISBN 0521 25705 0 Hardback GBPound 5.5
Tissues and Organs R.G. Kessel & R.H. Kardon W.H. Freeman S.F. ISBN 0-7167-0091-3 } }
Thanks to all who replied regarding my question on carbon contamination rates. It is really nice to be able to get the attention of so much expertise so easily. If anyone is interested I can send them a copy of all the replies I received, you can e-mail me at the address below. I plan to wait until my cold-finger gets installed (hopefully next week) and to try and find a place on campus to get the samples sputter cleaned (Getting them back to the lab will be another story :-)).
Hi I'm studying structure of emulsions of phospholipids, cholesterol, and triglycerides by fluorescence microscopy. I would appreciate hearing suggestions or comments about probes to use that might specifically label triglyceride droplets but not phospholipid membranes. I can work with a label added either before or after the emulsion is made.
For example I can specifically label phospholipid membranes using phospholipids with a fluorescent headgroup such as Bodipy-FL. I have used Nile Red (added either during or after manufacture of the emulsion) to simultaneously stain neutral lipids, but it also stains the membranes to some extent. It is also hard to work with because it photobleaches so quickly. Has anyone found useful alternatives to Nile Red for more-or-less specifically staining neutral lipid? Have you found a good additive that can minimize photobleaching of Nile Red?
Does DiI stain triglyceride droplets well, and would any related compound work better? Is NBD-cholesterol useful for imaging? (My impression is that it might photobleach rapidly.)
Thanks very much Richard Thrift Depotech Corp. Richard_Thrift-at-Depotech.com
Hi again I'm looking for a fluorescent probe that can demonstrate whether membranes are continuous or not. If I add the probe to the outer membrane I want it able to label internal membranes IF AND ONLY IF they are continuous with the external membrane. (In a dead or ATP-depleted cell, for instance. I'm actually studying liposomes.) It should be able to flip-flop and label the opposite leaflet of the bilayer to which it is exposed. (Actually, a probe that is unable to flip-flop would be useful, too, though not as useful.) It should spread only by diffusion in the "plane" of the membrane, once the carrier solvent dissipates.
Can you tell me whether DiI (and related dyes) flip-flop rapidly across membranes to label both sides of a bilayer in absence of ATP)? I've heard DiI labels all internal cell membranes; does it diffuse through cytoplasm or is it constrained to stay in the membrane? Can it jump from one membrane to another that is very closely apposed but not continuous? Is it a substrate for lipid transfer/exchange proteins, including albumin? Can you suggest any good references on properties of these dyes, or regarding their use for this particular purpose?
Pardon my ignorance; any suggestions would be greatly appreciated! THANKS very much ! Richard Thrift Depotech Corp. Richard_Thrift-at-Depotech.com (619) 625-2424
A friend of mine would like to obtain some prepared LM slides for teaching purposes (such as blood cells, onion skin, etc.). Any suggestions as to a relatively inexpensive source? TIA
-Bob ********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com **********************************
Hi, I work in biology with a STEM. I make tomographic reconstruction with a serial tiltes views, but I have a problem about realignement of projections because during the acquisition the rotation axis is shifted (5-10 pixels in X direction). Do you have a suggestion ???? Think you for your help.
Laurent Heliot
--
----------------------------**********----------------------------------- Laurent HELIOT LABORATOIRES DYOGEN/INFODIS (Post-doctorant) Institut Albert Bonniot, Mots Clefs : GRENOBLE Nucleolus, Nucleus, chromosome, Domaine de la MERCI AgNOR, Tomography, STEM, reconstruction 3D 38700 La TRONCHE ----------------------------------------------------------------------------
How about Edmund Scientific in NJ - call 609-573-6250 for a catalog.
Scott D. Ireland Media Cybernetics, LP " The Imaging Experts " scott-at-mediacy.com (716) 473-0222 Tel (716) 473-8048 Fax
---------- } From: Bob Citron {Bob_Citron-at-cc.chiron.com} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Prepared LM Slides } Date: Friday, October 11, 1996 7:01 PM } } Hi All; } } A friend of mine would like to obtain some prepared LM slides for teaching } purposes (such as blood cells, onion skin, etc.). Any suggestions as to a } relatively inexpensive source? TIA } } -Bob } ********************************** } Bob Citron } Chiron Vision } 555 W. Arrow Hwy } Claremont, CA 91711 } ph: (909)399-1311 } email: Bob_Citron-at-cc.chiron.com } **********************************
} Hi All; } } A friend of mine would like to obtain some prepared LM slides for teaching } purposes (such as blood cells, onion skin, etc.). Any suggestions as to a } relatively inexpensive source? TIA
Basics like that are carried by Carolina Biological Supply:800-334-5551. If this is for precollege education, consider the "do-it-yourself" approach rather than prepared slides - see the ProjectMICRO website. I'll be happy to suggest specific books.
Caroline Schooley Educational Outreach Coordinator, Microscopy Society of America Box 117 (45301 Caspar Point Road) Caspar, CA 95420 Phone/FAX: (707)964-9460 Email: schooley-at-mcn.org Web: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
If there are any Singaporean microscopists on the List who are willing to have a chat about work and general life in the region, would you please contact me direct on the email address below.
Thanks,
:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: Geoff Avern Manager Microscopy Laboratories Australian Museum Email: geoffa-at-amsg.austmus.gov.au 6 College St Ph: (61)(2) 9320 6198 Sydney, Australia. 2000 Fax: (61)(2) 9320 6059 ::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
A lateral response to your question: If your membranes include phosphatydilserine, you could try labelling them with Annexin, which is a short peptide that is specific for PS. It is available conjugated to fluorescein or biotin. It's used to study apoptosis, where PS, which is normally restricted to the inner lamella, flips over to the outer, allowing Annexin V to bind and label the cell.
Note that Annexin V isn't actually incorporated into the membrane, but binds strongly to components whose migration it would reveal. It only binds in the presence of fairly high calcium concentrations, and can be removed from membranes by washing in a calcium-free buffer.
Ray
VAt 2:30 pm 11/10/96, Richard Thrift wrote: } Hi again } I'm looking for a fluorescent probe that can demonstrate whether } membranes are continuous or not. If I add the probe to the outer } membrane I want it able to label internal membranes IF AND ONLY IF } they are continuous with the external membrane. (In a dead or } ATP-depleted cell, for instance. I'm actually studying liposomes.) It } should be able to flip-flop and label the opposite leaflet of the bilayer to } which it is exposed. (Actually, a probe that is unable to flip-flop would } be useful, too, though not as useful.) It should spread only by diffusion } in the "plane" of the membrane, once the carrier solvent dissipates. } } Can you tell me whether DiI (and related dyes) flip-flop rapidly across } membranes to label both sides of a bilayer in absence of ATP)? I've } heard DiI labels all internal cell membranes; does it diffuse through } cytoplasm or is it constrained to stay in the membrane? Can it jump } from one membrane to another that is very closely apposed but not } continuous? Is it a substrate for lipid transfer/exchange proteins, } including albumin? Can you suggest any good references on properties } of these dyes, or regarding their use for this particular purpose? } } Pardon my ignorance; any suggestions would be greatly appreciated! } THANKS very much ! } Richard Thrift } Depotech Corp. } Richard_Thrift-at-Depotech.com } (619) 625-2424
Just wanted to thank everyone who replied so quickly to my question about prepared microscope slides. The most highly recommended sources included Triarch (Ripon, WI) and Carolina Biological Supply (800) 334-5551.
-Bob ******************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 (909)399-1311 email: Bob_Citron-at-cc.chiron.com ********************************
Another catalog source is NASCO. Their scientific catalog has many more slides of basic biological specimens than Edmund's catalog. I took the following information is from NASCO's web site http://www.nascofa.com/ (You can order a catalog via the web site.) ______ Phone: Call Toll Free - U.S.A. & Canada 1-800-558-9595 If ordering from outside the U.S.A., call 1-414-563-2446
NASCO ? FORT ATKINSON 901 Janesville Avenue P.O. Box 901 Fort Atkinson, WI 53538-0901 E-mail: info-at-nascofa.com Fax: 414-563-8296
PHONE ORDER HOURS: Weekdays 7:00 AM - 6:00 PM Saturday 8:00 AM - 12:00 Noon Fort Atkinson, WI (CT) Modesto, CA (PT)
} } } Scott D. Ireland {sireland-at-frontiernet.net} 10/12/96 03:38 } } } How about Edmund Scientific in NJ - call 609-573-6250 for a catalog.
Scott D. Ireland Media Cybernetics, LP " The Imaging Experts " scott-at-mediacy.com (716) 473-0222 Tel (716) 473-8048 Fax
---------- } From: Bob Citron {Bob_Citron-at-cc.chiron.com} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Prepared LM Slides } Date: Friday, October 11, 1996 7:01 PM } } Hi All; } } A friend of mine would like to obtain some prepared LM slides for teaching } purposes (such as blood cells, onion skin, etc.). Any suggestions as to a } relatively inexpensive source? TIA } } -Bob } ********************************** } Bob Citron } Chiron Vision } 555 W. Arrow Hwy } Claremont, CA 91711 } ph: (909)399-1311 } email: Bob_Citron-at-cc.chiron.com } **********************************
issued by OSRAM GmbH, Photo-Optics Marketing, Berlin Fax: (Germany)-30 -3386 -2773, e-mail: osramwg-at-aol.com
re: HBO 50 W/AC, lamp for microscopes:start up..
Dear subscribers,
this is now the third statement, related to "start-up time".
Some of you complained about the startup time in HBO 50 W/AC lamps.
Beginning of September, we have started delivery of a number of slightly modified lamps. The obvious difference is, that the lamps have a reflector on both sides of the electrodes (standard lamp has only one reflector). These lamps are 100% compatible to the standard lamps, however we found a reduced start-up time. It will be important for us to get your feedback. If you find your next lamp with reflectors on both sides we would like to ask you to give us some kind of feedback. This should help you and us as well by an increased performance of the lamps in the future.
Thank you very much for your support.
Statement NR. 4 will target "low output power from the lamp"
I would be grateful if you could let postdoctoral people in your department know that there was an advertisement for a Marine Protistan Biologist in the Antarctic Division in the WEEKEND AUSTRALIAN on 12 October.
Cheers
Harvey
********************************************************************* Harvey J. Marchant Program Leader, Biology Australian Antarctic Division Channel Highway, Kingston, Tasmania 7050 Australia
issued by OSRAM GmbH, Photo-Optics Marketing, Berlin Fax: (Germany)-30 -3386 -2773, e-mail: osramwg-at-aol.com
re: HBO 50 W/AC, lamp for microscopes, no or difficult ignition
Dear subscribers,
this is not a commercial advertisement but (hopefully) valuable information to all microscopists, who recently faced problems with OSRAM lamps in their microscopes. The first statement is related to " no starters or difficult ignition". End of 1995, we have replaced a machine in production. This new machine had at the beginning a broader tolerance regarding fill pressure and therefore a small number of lamps have been produced with a wrong fill pressure, causing ignition problems. However it took several month until we heard about these problems and until the failure was located. Corrective actions have been taken and the problem should not exist any longer. However, dependent on the stock situation of the different microscope manufacturers, service organizations and lamp dealers, there are probably still a certain number of these lamps in the field. If you do have any problem with ignition AND your lamp shows the following serial numbers: xxxx qP2....xxxxqP6 or xxxxqk1.... qk4, please report to your dealer or to us directly. Your lamps will be replaced free of charge.
Statement NR. 2 will follow soon and contains information about reported lamp explosions
issued by OSRAM GmbH, Photo-Optics Marketing, Berlin Fax: (Germany)-30 -3386 -2773, e-mail: osramwg-at-aol.com
re: HBO 50 W/AC, HBO 100 W/2 and HBO 103W/2, lamp for microscopes: low output power on Nikon microscopes
Dear subscribers,
this is now statement NR. 4, related to low light output.
Two reasons in general might cause low output: - blackening of the bulb (aging) - low current from the psu
Blackening is dependent on the voltage, current (also power supply!), type of current (DC vs., AC) electrode material, gas purity etc... etc. With HBO 50 W/AC it is an aging process.
Low output from the microscope with HBO 100 W/2 or HBO 103 W/2 it is mostly related to the characteristics of the power supply. An HBO lamp has a burning voltage depending on the lamp itself and draws as much current as it can get. The power supply however powers the lamp depending on the lamp voltage. For Nikon microscopes for example, the power to the lamp will be between 60 and 100 W depending on the lamp voltage (which increases as a function of lifetime- if you are interested in this graph- please send an e-mail, we will get it to you immediately). Other microscope manufacturers have decided for constant power mode. In this case, the lamp demands increasing current over lifetime and you will get a more or less constant output until the end of lifetime.
If you do have any problem with Nikon equipment regarding low output power, we recommend to contact your Nikon dealer. We were told today from NIKON Yokohama, that there is a problem with their power supplies, however up to now, we do not have more detailed information.
The last statement NR. 5 will summarize faq's from the microscopy list subscribers during the last weeks
issued by OSRAM GmbH, Photo-Optics Marketing, Berlin Fax: (Germany)-30 -3386 -2773, e-mail: osramwg-at-aol.com
re: HBO 50 W/AC, HBO 100 W/2 and HBO 103 W/2, lamp for microscopes: lamp explosions
Dear subscribers,
this is now the second statement, related to "lamp explosions"
A very small number of you (6 people) reported, that they have been faced with lamp explosions in their microscopes. We have investigated this situation here in the company, however within the last years (100.000 produced lamps) no explosions have been reported. The questions was: When does a lamp explode (beside operation above the rated lifetime)?-either due to a defect in the bulb (microcrack etc.) or due to overheating in the lamp (by excessive energy input, by backreflection of light into the bulb or by improper cooling) or due to use above rated lifetime. Therefore we got back to the microscope suppliers and this is what we found: Explosions have been reported on Zeiss microscopes. In old equipment, lamp explosion might be caused by a slightly misaligned optical system, i.e. when the backreflected light (from the rear mirror) is going back directly into the electrode area of the lamp. This might cause overheating of the lamp with a reduced lifetime or explosion. In Nikon microscope: We did some testing on lamp housing from Nikon with HBO 103W/2. Here we found, that in some cases the measured temperature was close to the upper level of the tolerated temperature, but still within the given specs. From users it was also reported, that contact between lamp and lamp holder might create problems in a very few cases due to improper fitting. This might result in local overheating ( the metal on the lamp base gets a bluish color) and cracking the lamp. The cracking of the lamp might be proceeded by a flickering, caused by arcing between lamp and lamp holder. If you find your bases with a blue color, please report to your Nikon dealer or service for inspection of the lamp holder. Under proper contact conditions, no explosion will happen.
Statement NR. 3 will follow soon and contains information about start up time.
issued by OSRAM GmbH, Photo-Optics Marketing, Berlin Fax: (Germany)-30 -3386 -2773, e-mail: osramwg-at-aol.com
re: HBO 50 W/AC, HBO 100 W/2 and HBO 103 W/2, lamp for microscopes: FAQ's
Dear subscribers,
this is now the last statement NR. 5, related to frequently asked questions.
1.) What are the technical date for my lamp? Please send an e-mail or a fax with your address- we will get you the latest datasheets.
2.) Can I replace my HBO 100/2 by the longlife version HBO 103/2? In principle...You can however.... The electrical characteristics are about the same. In addition, we have tested all lamps in Zeiss, Nikon etc. equipment available on the market. No problems have been found or reported from customers so far. However... - for old Nikon equipment we currently recommend the HBO 100W/2 since there is a better coupling between power supply and lamp due to the lower lamp voltage. - for Zeiss equipment: please ask your Zeiss service about the optical design in your particular lamp house. Zeiss has changed their optical setup a certain time ago- new equipment therefore should not see any problems.
3.) My HBO 50 W/AC failed after about 50 h to ignite (cool room due to air condition) , I used a hair dryer, warmed up the lamp and than it started again- does this damage the lamp? The hair dryer either evaporates a small portion of the mercury from the surface of the electrode inside the lamp and makes it easier for the cold lamp to start. Or it heats up the bulb connections (see statement NR. 3) A moderate heating will not damage the lamp ( I do not know what happens to the lamp housing and the paint.), however this should not be the way for a microscopist to work with OSRAM lamps. I assume, that this was either one of the lamps, mentioned in statement NR. 1 or it was a contact problem like we have seen it from Nikon equipment). In the first case, report to your OSRAM dealer or to us directly (include Serial number and type of microscope), we will replace your lamp. In the second case, you should report to your microscope dealer.
4.) Can I install an additional fan to reduce risk of explosion and to increase lifetime? We do not recommend to do this because: - under normal circumstances you should not have an explosion, please refer to previous statement NR. 2 about possible reasons for an explosion. - cooling of the lamp will reduce output power due to condensation of the mercury - cooling of the lamp might create tensions in the bulb and cause and explosion
5.) If my lamp (HBO 50 W/AC) in my ZEISS microscope does not ignite, I was told to press a special button. This "special button" is the reset button for the igniter. The starter disables after a certain number of unsuccessful attempts to ignite the lamp or when the current has exceeded a certain value. If there is no ignition, please press this little button. Refer to the Zeiss users manual!
6.) My microscope service told me, that according the newest research there is absolutely no reason for the two images not to be aligned after lamp replacement. In general, this is not correct and might damage your system. Please refer to your handbook or ask your microscope supplier about the proper alignment.
This was the last statement on the recently reported problems on HBO lamps for microscopes. Please feel free to contact us directly in case you have any further questions: osramwg-at-aol.com
I am a Student at the University of Lethbridge and I am taking a Microscopy course related to the study of Microcircuitry. My professor is a biological Microscopist and has rather limited experience with the study of circuitry. As this is the case I was wondering if anyone on this list has suggestions for things to study related to this and any experiments that might be interesting.
I need to quantify pavements, which can be looked at as a system of packed particles. I need to obtain the number of contacts each particle has on average with another particle. This can then be related to mechanical properties such as shear resistance, etc.
Are there microscopic techniques to get this information from a compact and stereology methods to extract such information. Any response will be appreciated.
My name is Nancy Zjaba and I am a student in the Madison (Wis.) Area Technical College EM program. As part of my coursework, I am required to give an oral report. My topic is the use of EM in forensic science. I have done preliminary research, but have specific questions I'm hoping someone will help me with (or point me to a reference):
1. When did SEM and TEM first pass the Frye rule as acceptable techniques for examining evidence?
2. How is it that the courts have limited the kind of sample preparation that can be done? What common preparation techniques can be used, and which ones can't?
3. Are people who conduct forensic EM required to testify in court? How often? Does maintaining the "chain of evidence" require a lot of time (as far as paperwork)?
We were cleaning a closet when we found a brandy new (we think) CRT for a Cambridge 600. (host instrument retired here in 1990) Specs on the box read as follows: Thorn Brimar limited, Type M17/151BE, serial No. 430142. Why let you know? Well, it's available....for free!
Now, not having shipped one of these babies before, I note that there is a cautionary note on the (original) packing about the contents being: "glass", "evacuated tube", etc. Do any of you know if there are any restrictions on mailing a CRT?
Qualifications for donation: not exactly a first come - first serve...rather, a first "needy" come - first serve. Please mail me with your requests (if there be any at all) and I will choose a winner.
Winton Cornell
P.S. I will need a FedEx # or like for the mailing
Dr. Winton Cornell Senior Research Associate Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
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At 02:00 PM 10/15/96 -0500, Nancy Zjaba wrote: } My name is Nancy Zjaba and I am a student in the Madison (Wis.) Area } Technical College EM program. As part of my coursework, I am required to } give an oral report. My topic is the use of EM in forensic science. ...
This is an interesting topic ... please reply to the list ... TIA.
cheers, shaf
{\/} /\ {\/} /\ {\/} /\ {\/} cognito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
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I am considering purchasing a negative film scanner and I have narrowed the choices to the Polaroid Sprintscan 45, the Nikon LS4500AF or the LeafScan 45. The LeafScan 45 is no longer produced but I may be able to get a hold of one of the last ones or a rebuilt one. I have used the LeafScan 45, but I have no experience with either the Polaroid or the Nikon. The LeafScan exceeds the performance of the other two (according to the specifications), but is the cost justified? The Polaroid and Nikon are approximately half the price of the LeafScan.
I would appreciate hearing your comments (advantages/disadvantages) if you have used either the Polaroid or the Nikon, especially if you have experience with the LeafScan for comparison purposes.
Thanks in advance. ******************************************************* David F. Teter Los Alamos National Laboratory Materials Science and Technology: Metallurgy (MST-6) Mail Stop: G755 Los Alamos, NM 87545 ph: (505)665-0160 fax: (505) 665-0657 e-mail: teter-at-lanl.gov *******************************************************
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At 12:40 15/10/96 -0600, you wrote: } } I am considering purchasing a negative film scanner and I have narrowed the } choices to the Polaroid Sprintscan 45, the Nikon LS4500AF or the LeafScan } 45. The LeafScan 45 is no longer produced but I may be able to get a hold of } one of the last ones or a rebuilt one. I have used the LeafScan 45, but I } have no experience with either the Polaroid or the Nikon. The LeafScan } exceeds the performance of the other two (according to the specifications), } but is the cost justified? The Polaroid and Nikon are approximately half the } price of the LeafScan. } } I would appreciate hearing your comments (advantages/disadvantages) if you } have used either the Polaroid or the Nikon, especially if you have } experience with the LeafScan for comparison purposes. } Thanks in advance. } ******************************************************* } David F. Teter } Los Alamos National Laboratory } Materials Science and Technology: Metallurgy (MST-6) } Mail Stop: G755 } Los Alamos, NM 87545 } ph: (505)665-0160 fax: (505) 665-0657 } e-mail: teter-at-lanl.gov } ******************************************************* David & all: I have an interest in digital cameras - we sell the MicroLumina in Australasia. However, I truly believe that a high resolution digital camera, although more expensive, is a better solution than a scanner. I don't like scanners, I've had too much trouble with scan lines etc. Combined with a macro lens a digital camera can copy negs on a light box beautifully, it can also "copy" prints, apparatus, people and record microscope images. In short it's infinitely more versatile. Jim Darley
Probing & Structure } (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } A great microscopy site http://www.ultra.net.au/~pns/
Hello All, We are having a problem that I wondered if anyone has had experience with out there. We actually have two problems that may or may not be associated. We have been getting up to 5% Sr detected when doing quantitative analysis with specimens that are high in Si (~20%) where no peaks for Sr are detected on the spectra (even at 35kv) and there should be no Sr present in the sample. The samples are charging at times due to the high Si but this is not always the case. We are concerned about the proximity of the peaks causing some type of overlap for some reason (Si and Sr)
We are also getting count rate variations with alterations in contrast and brightness settings while in both back scatter and secondary modes respectively. These alterations are not having an effect on our total area readings but they do influence the count rate and dead time.
We have tried conditioning the analyzer, multiple calibrations, changing electrical outlets for the back scatter detector, carbon coating the specimens... We also noted recently that our backscatter detector had slipped and was almost touching the secondary detector though this has been corrected and is no longer a problem. Analysis of copper/gold standards are giving good quantitative results.
I realize that these details are somewhat sketchy but does anyone recognize this problem? Any input is greatly appreciated.
Wayne England ORTECH CORP. Mississauga, Ontario wengland-at-ortech.on.ca
I like to know how I can make a relay lenssystem for connecting a CCD camera to the camera port of a lightmicroscope. I want to see 1/3 to 3/4 of the visual field on the monitor.
I got a "Stump The Prof" question today. If formaldehyde is a monofunctional fixative, how can it cross-link anything and, by extension, how can it be a fixative at all?
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
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Referring to Winton Cornell's offer of a free M17/151BE CRT, VG HB5/HB501 owners may like to check on the part numbers of their CRT's - I've a feeling they are the same. But then, my memory may fail me!
Tony Garratt-Reed
********************************* ** ** ** Anthony J. Garratt-Reed ** ** MIT Room 13-1027 ** ** 77 Massachusetts Avenue ** ** Cambridge, MA 02139-4307 ** ** USA ** ** ** ** Ph: 617-253-4622 ** ** Fax: 617-258-6478 ** ** ** ********************************* *********************************
Message-Id: {199610161349.IAA05368-at-Sparc5.Microscopy.Com} To: Microscopy messageslistserver {Microscopy-at-Sparc5.Microscopy.Com}
} I got a "Stump The Prof" question today. If formaldehyde is a } monofunctional fixative, how can it cross-link anything and, by extension, } how can it be a fixative at all? } } Bob
From J.A. Kiernan, _Histological & Histochemical Methods_, pg 18ff:
formaldehyde reacts with water to form methylene hydrate HOCH2OH this then reacts with various parts of proteins to form methylene cross-links. But I've also heard that it works by a two-stage reaction, where 2 CHO's react to form some unspecified product with two functional groups which then cross-link the proteins. I prefer the first explanation. Phil
&&&&&&&&&&& Illigitmi non carborundum &&&&&&&&&&&
Philip Oshel *all mail:* University of Illinois Station A Rm 74 Bevier Hall PO Box 5037 905 S. Goodwin Ave. Champaign, IL 61825-5037 USA Urbana, IL 61801 (217) 244-3145 oshel-at-ux1.cso.uiuc.edu (217) 355-1143 home
This puzzled me once also but I think I now understand.
RH + CH2O {=} RCH2(OH)
Then
RCH2(OH) + HR' {=} RCH2R' + H2O
note that this is a reversible reaction so with extensive washing one can lose the fixation. could this be how some "antigen retrival" processes work?
Most of the formaldehyde in aqueous solution forms a glycol called methylene glycol. the small formaldehyde penetrates the tissue rapidly. as the formaldehyde reacts with tissue, it shifts the equilibrium and more methylene glycol turns into formaldehyde. this latter transition is one reason formaldehyde is a slow fixer despite being a rapid pentetrator.
Also with time, polymers of formaldehyde form and can substitute directly into this equation. This is why older stocks of formalin or formaldehyde can act differently from fresh stocks.
Hope this helps. Tom Phillips.
} I got a "Stump The Prof" question today. If formaldehyde is a } monofunctional fixative, how can it cross-link anything and, by extension, } how can it be a fixative at all? } } Bob } } } Robert R. Wise, PhD } Director, UWO Electron Microscope Facility } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-uwosh.edu
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Department of Materials Science and Engineering & Laboratory for Research on the Structure of Matter 3231 Walnut Street University of Pennsylvania Philadelphia, PA 19104-6272
Qualified candidates will have a Ph.D. in a physical science or engineering field as well as considerable experience with transmission electron microscopy. Preferred candidates will have experience with beam sensitive materials, EELS, and/or polymer microscopy. The successful candidate will initiate two or more new projects associated with (1) lateral uniformity at polymer interfaces, (2) electrically active polymeric devices, and/or (3) optically active polymers. These projects will involve the (soon to arrive) field emission analytical TEM.
Start Date is flexible. Earliest start date: December 1, 1996 Latest start date: April 1, 1996
To apply please send the following items to the address above: 1. complete curriculum vitae 2. names and addresses of two references 3. one preprints/reprints
Applications are due November 20, 1996.
regards, Prof. Karen I. Winey winey-at-lrsm.upenn.edu
----------------------------------------------------------------------------- Karen I. Winey Assistant Professor (215) 898-0593 Materials Science and Engineering Department (215) 573-2128 FAX 3231 Walnut Street winey-at-lrsm.upenn.edu University of Pennsylvania LRSM building; room 308 Philadelphia, PA 19104-6272
Department of Materials Science and Engineering & Laboratory for Research on the Structure of Matter 3231 Walnut Street University of Pennsylvania Philadelphia, PA 19104-6272
Qualified candidates will have a Ph.D. in a physical science or engineering field as well as considerable experience with transmission electron microscopy. Preferred candidates will have experience with beam sensitive materials, EELS, and/or polymer microscopy. The successful candidate will initiate two or more new projects associated with (1) lateral uniformity at polymer interfaces, (2) electrically active polymeric devices, and/or (3) optically active polymers. These projects will involve the (soon to arrive) field emission analytical TEM.
Start Date is flexible. Earliest start date: December 1, 1996 Latest start date: April 1, 1996
To apply please send the following items to the address above: 1. complete curriculum vitae 2. names and addresses of two references 3. one preprints/reprints
Applications are due November 20, 1996.
regards, Prof. Karen I. Winey winey-at-lrsm.upenn.edu
----------------------------------------------------------------------------- Karen I. Winey Assistant Professor (215) 898-0593 Materials Science and Engineering Department (215) 573-2128 FAX 3231 Walnut Street winey-at-lrsm.upenn.edu University of Pennsylvania LRSM building; room 308 Philadelphia, PA 19104-6272
Why is Sr even showing up in your analyses? I know we can help or hurt our standardless results by confusing the element list. I would rather not let the analyzer try to figure out which elements are present, or if it tries to revise the list to the proper elements. (I got very tired of always deleting V from the list of identified elements when O was present.) Without standards, a certain amount of Sr may help the fit to your Si peak. It depends on how good a shape they use. This is another arguement for using standards whenever possible - both a proper peak shape and a real intensity reference.
Regarding the fluctuating count rate - Are you running a "shotgun" analysis over an area at a "slow" scan rate on a heterogenous sample? We set the count rate for x-ray maps on concrete and watch the count rate wander 10% and more as the beam scans the field. We are now setting it using TV scan rate, although it doesn't matter a lot except for consistency.
At 08:53 AM 10/16/96 -0400, you wrote: } } Hello All, } We are having a problem that I wondered if anyone has had experience with } out there. } We actually have two problems that may or may not be associated. We have } been getting up to 5% Sr detected when doing quantitative analysis with } specimens that are high in Si (~20%) where no peaks for Sr are detected on } the spectra (even at 35kv) and there should be no Sr present in the sample. } The samples are charging at times due to the high Si but this is not always } the case. We are concerned about the proximity of the peaks causing some } type of overlap for some reason (Si and Sr) } } We are also getting count rate variations with alterations in contrast and } brightness settings while in both back scatter and secondary modes } respectively. These alterations are not having an effect on our total area } readings but they do influence the count rate and dead time. } } We have tried conditioning the analyzer, multiple calibrations, changing } electrical outlets for the back scatter detector, carbon coating the } specimens... We also noted recently that our backscatter detector had } slipped and was almost touching the secondary detector though this has been } corrected and is no longer a problem. } Analysis of copper/gold standards are giving good quantitative results. } } I realize that these details are somewhat sketchy but does anyone recognize } this problem? Any input is greatly appreciated. } } Wayne England ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
This classical paper may interest you also: "Current efforts to attain the resolution limit of the transmission electron microscope" by E. Ruska, Journal of the Royal Microscopical Society, Vol. 84, Pt. 1, April 1965. Pp. 77-103.
to Pieter Houpt, who asks about relay optics to connect a c-mount camera to a camera port:
We have mounted various cameras on the ports of many microscopes, and the general conclusion is that they are all different. The usual relay optic is used to step the real image from the port down to the size of the camera sensor. The details of a design depend on the sensor size and the position of the image when the microscope is in focus. I have seen systems as simple as a singlet lens (not very satisfactory) to a singlet and doublet combination (worked well). To a large extent, the difficulties are mechanical.
We actually sell a relay lens that fits into a DIN 23mm eyepiece hole, with or without a clamp assembly, and has a c-mount thread on the other end. We use it with many of our systems. If you would like more info, Email me.
I can also give you the name of a company that makes camera adapters for most of the standard light microscopes.
} I got a "Stump The Prof" question today. If formaldehyde is a } monofunctional fixative, how can it cross-link anything and, by extension, } how can it be a fixative at all? } Isn't formaldehyde cross-linked to itself to some extent - i.e. once made up into solution it's in small polymers not pure monomers? I think this is the case even for solutions made from paraformaldehyde powder. And the C in HCHO should be able to cross-link two Ns from different amino groups or peptide links. Acrolein is also a monoaldehyde, I think, but is a very efficient fixative so must work in a similar way. I'm not 100% sure about this, and am no chemist, just remember it from long ago EM prep classes.
Rosemary
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Clayton, Victoria 3168, Australia phone 61-3-9905 5670 email r.g.white-at-sci.monash.edu.au fax 61-3-9905 5613 or rgwhite-at-vaxc.cc.monash.edu.au
} } I am considering purchasing a negative film scanner and I have narrowed the } choices to the Polaroid Sprintscan 45, the Nikon LS4500AF or the LeafScan } 45. David, if you have the money, I will definitely recommend you purchasing the LeafScan 45. I have been using the LeafScan 45 for the last two years, and I'm very happy with it. The resolution and reliability of the scan are more superior than the Polaroid Sprintscan, and the Nikon LS4500AF.
--Ciprian
_________________________________________________________ Ciprian A. Almonte Rochester Institute of Technology Biomedical Photographic Communications Rochester, NY 14623-5603
Visit my web site at http://www.isc.rit.edu/~caa3045/ __________________________________________________________
For a recent book, which explains this and many other problems in EM, check the monograph Fine Structure Immunocytochemistry (ed: Gareth Griffiths, EMBL) Springer, Berlin, 1993 (ISBN 3 540 54805 X) or New York (ISBN 0 387 54805 X) and many references cited therein. (pp. 39 ff on formaldehyde fixation) Reinhard Rachel
Dear Winton; Your message of Tuesday October is rather intriguing. You had written, and I quote from your message "We were cleaning a closet when we found a brandy----". I'm sure many of us as we read our e-mail at unsocial hours would have had a sharp focus of interest which may have dropped away as the message continued on about CRT etc. I hope you find a home for your CRT.
Patrick Echlin
On Tue, 15 Oct 1996, Winton Cornell wrote:
} Scopers: } } We were cleaning a closet when we found a brandy new (we think) CRT for a } Cambridge 600. (host instrument retired here in 1990) Specs on the box } read as follows: Thorn Brimar limited, Type M17/151BE, serial No. 430142. } Why let you know? Well, it's available....for free! } } Now, not having shipped one of these babies before, I note that there is a } cautionary note on the (original) packing about the contents being: } "glass", "evacuated tube", etc. Do any of you know if there are any } restrictions on mailing a CRT? } } Qualifications for donation: not exactly a first come - first } serve...rather, a first "needy" come - first serve. Please mail me with } your requests (if there be any at all) and I will choose a winner. } } Winton Cornell } } P.S. I will need a FedEx # or like for the mailing } } } Dr. Winton Cornell } Senior Research Associate } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu } } }
Charging will affect SE much more than BSE, but I have certainly had specimens charge to the point of disrupting BSE imaging. Charging will also reduce the effective incident beam potential, causing possible loss of higher energy x-ray counts. This will be evident when examining the x-ray background signal. With no charging the bkgn should "tail-off" at the beam potential. As a specimen charges, the maximum bkgn energy will become lower. For example, an incident beam of 10kV will normally produce a background terminating at 10kV. If the specimen has charged to 2kV, the bkgn will terminate at (10-2) 8kV. Woody ______________________________ Reply Separator _________________________________ Subject: count rate fluctuations
The samples are charging at times ...SNIP...
We are also getting count rate variations with alterations in contrast and brightness settings while in both back scatter and secondary modes respectively. ...SNIP...
Wayne England ORTECH CORP. Mississauga, Ontario wengland-at-ortech.on.ca
The Minnesota Microscopy Society is involved in a preliminary trial of the Project Micro Program. One of the experiments calls for examining various sands at low magnification (10x-20x). These sands should have unique morphological or color properties which could be seen at this magnification. If anyone has access to some unique sand we would appreciate the help. Please contact me at my EMail address
I am thinking about buying a used Hummer V Sputter Coater. To those unfamiliar with the model, it is a completely self-contained unit (i.e., it has an integral vacuum pump) and is also capable of carbon coating. The instrument is about 8 years old, but has hardly seen any use. It basically looks brand new and comes with a gold-palladium source and a carbon source. It has been cleaned, greased, oiled, etc., and it works well. I have two questions:
1. Is this a good, reliable, instrument?
2. Is $2500 a fair asking price.
Thanks.
Martin A. Levin Department of Biology Eastern Connecticut State University Willimantic, CT 06226 Phone: (860)465-4324 FAX: (860)465-5213
Wayne: It is not clear if you are using WDS or EDS or which lines you are = measuring but regardless, the Si K-beta line is close enough to Sr = L-alpha to cause spurious counts for Sr, even with the better-resolving = WDS technique. My experience on a microprobe with wavelength = spectrometers is that Sr measurement below ~ 0.5 wt% SrO in a silicate = with ~50 wt% SiO2 was quite difficult unless some overlap corrections = are made. =20 Your description of count rate variations in BSE or SE mode is = difficult to understand, but if you are collecting element intensity = data while scanning a heterogeneous target, of course you will get = fluctuations. If Si varies in your sample, then the Sr data due to Si = overlap will also vary. Did I understand your problem correctly? -Jim McGee *.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.* James J. McGee (jmcgee-at-sc.edu) Dept. of Geological Sciences University of South Carolina (803) 777-6300 (Office) Columbia, SC 29208 (803) 777-6610 (Fax)
----------
Hello All, We are having a problem that I wondered if anyone has had experience = with=20 out there. We actually have two problems that may or may not be associated. We = have=20 been getting up to 5% Sr detected when doing quantitative analysis with=20 specimens that are high in Si (~20%) where no peaks for Sr are detected = on=20 the spectra (even at 35kv) and there should be no Sr present in the = sample.=20 The samples are charging at times due to the high Si but this is not = always=20 the case. We are concerned about the proximity of the peaks causing = some=20 type of overlap for some reason (Si and Sr)
We are also getting count rate variations with alterations in contrast = and=20 brightness settings while in both back scatter and secondary modes=20 respectively. These alterations are not having an effect on our total = area=20 readings but they do influence the count rate and dead time. ...
Wayne England ORTECH CORP. Mississauga, Ontario wengland-at-ortech.on.ca
I was wondering about the kinds of BSE detector systems that people have with their SEM's. What are your opinions? How reliable is the detector system that you have? Which one/one's would you recommend?
Thanks in advance, Peling Melville
-------------------------------------------------------------- Peling Fong Melville Senior Scientific Assistant Interdepartmental Facilities American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 U.S.A. ****************************** E-mail: peling-at-amnh.org Work #: (212) 769-5469 FAX #: (212) 769-5495
On of the researchers here is doing toxicologic research on marine mammmal specimens collected in a field situation. Specifically, she is concerned with the localization of heavy metal contaminants in tissues at a cellular and subcellular level. Tissues will be analyzed for metal concentrations via atomic absorption spectroscopy (AA). In addition, autometallographic silver development of tissue slide preparations at the light and electron microscopic level will be used to both amplify and localize metal deposition in tissue. However, she is looking for a technique that would positively identify minute amounts of metals in slide preparations that would serve as an adjunct to autometallographic and AA results. The elements of particular interest are mercury, silver, and selenium. I am anticipating that concentrations in tissues will range from limits of detection (for AA) to possibly a few hundred ppm, with most concentrations being in the 5-50 ppm range, thus very likely to be outside usual EDX limits. Is PIXE a useful adjunct technique? Any advice with respect to a technique designed to identify these elements at low concentration in situ would be greatly appreciated.
************************ Richard Crang Dept. Plant Biology University Illinois Urbana, IL 61801 (217) 244-3143 ************************
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
We've been using a Hummer IV for about 14 years, and it's done very well. There were vacuum leak problems, but we sorted them out. Price? No idea, sorry.
Would you be interested in sand collected by my well pump filter? The grains are sharp edged and irregular - about 100 microns long. The sand is predominantly an even mix of quartz (clear/white) and nearly black particles. I forget the composition of the black - I think it was comprised of O, Si, Fe, and maybe(?): Mg, Ca, Mn, and or Ti?? A geologist friend belives it comes from decomposing (poorly "sintered") igneous(sp?) rock which was once an undersea deposit of volcanic ash.
Woody White Electron Microscopist Wk: woody.n.white-at-mcdermott.com Hm: woody.white-at-worldnet.att.net http://www.geocities.com/capecanaveral/3722
The Minnesota Microscopy Society is involved in a preliminary trial of the Project Micro Program. One of the experiments calls for examining various sands at low magnification (10x-20x). These sands should have unique morphological or color properties which could be seen at this magnification. If anyone has access to some unique sand we would appreciate the help. Please contact me at my EMail address
University of Illinois at Urbana-Champaign Materials Research Laboratory
RESEARCH ELECTRON MICROSCOPIST
The Materials Research Laboratory at the University of Illinois is seeking an experienced electron microscopist as a staff member in the Center for Microanalysis of Materials. The Center is a major research facility with eight electron microscopes as well as instruments in surface microanalysis, x-ray diffraction, and other analytical techniques.
The person will work mainly on SEM, but should have the experience and flexibility to work also in TEM and possibly other techniques, if needed. The Center has two SEMs and a third may be acquired shortly. Experience in one or more of the following techniques would be especially important: Cathodo-luminescence, EBSP, quantitative EDX. The main responsibilities of the position are to facilitate the research of the approximate 150 SEM users yearly, train new users, and keep the instruments running.
There will be ample opportunity to carry out interactive research in the facility with the wide range of research programs. Possibilities for independent research could also be developed.
This position requires a university degree in physics, materials science or a related field (PhD is desired) and at least three years experience with electron microscopes.
This is a 12 month, 100% time regular appointment with standard university benefits. Salary will be commensurate with education and experience. The position will be available in early 1997. In order to ensure full consideration, application must be received by November 30, 1996. Please send letter of application, resume, and names and addresses of three references to J. A. Eades, c/o Donna Jacobs, University of Illinois, Materials Research Laboratory, 104 South Goodwin Avenue, Urbana, Illinois 61801, phone (217) 244-2944. For technical information, call J. A. Eades at (217) 333-8396.
The University of Illinois is an Affirmative Action/Equal Opportunity Employer.
A colleague would like to study the development of polysaccharide biofilms (secreted by Pseudomonad bacteria) on lettuce leaves. She asked whether there were any quick and easy (or, if necessary, long and tricky) ways of seeing whether these polysaccharides were present on leaves before and after they are turned into salads. My suggestion was to use a simple polysaccharide stain on hand sections of leaves, but the film may be too thin to distinguish it from the cuticle, even if she compared leaves from sterile cultures with field-grown leaves. Any suggestions?
TIA,
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Clayton, Victoria 3168, Australia phone 61-3-9905 5670 email r.g.white-at-sci.monash.edu.au fax 61-3-9905 5613 or rgwhite-at-vaxc.cc.monash.edu.au
IMHO there's only one choice, the Robinson BSE detector. We do approx. 90% of our micrographs using ours and it's worked wonderfully for 10 years. The decision becomes 'which model of Robinson to get'?
Cheers,
:::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: Geoff Avern Manager Microscopy Laboratories Australian Museum Email: geoffa-at-amsg.austmus.oz.au 6 College St Ph: (61)(2) 9320 6198 Sydney, Australia. 2000 Fax: (61)(2) 9320 6059 :::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::: ______________________________ Reply Separator _________________________________
Dear Microscopists,
I was wondering about the kinds of BSE detector systems that people have with their SEM's. What are your opinions? How reliable is the detector system that you have? Which one/one's would you recommend?
Thanks in advance, Peling Melville
-------------------------------------------------------------- Peling Fong Melville Senior Scientific Assistant Interdepartmental Facilities American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 U.S.A. ****************************** E-mail: peling-at-amnh.org Work #: (212) 769-5469 FAX #: (212) 769-5495
} Hello All, } We are having a problem that I wondered if anyone has had experience with } out there. } We actually have two problems that may or may not be associated. We have } been getting up to 5% Sr detected when doing quantitative analysis with } specimens that are high in Si (~20%) where no peaks for Sr are detected on } the spectra (even at 35kv) and there should be no Sr present in the sample. } The samples are charging at times due to the high Si but this is not always } the case. We are concerned about the proximity of the peaks causing some } type of overlap for some reason (Si and Sr) } } We are also getting count rate variations with alterations in contrast and } brightness settings while in both back scatter and secondary modes } respectively. These alterations are not having an effect on our total area } readings but they do influence the count rate and dead time. } } We have tried conditioning the analyzer, multiple calibrations, changing } electrical outlets for the back scatter detector, carbon coating the } specimens... We also noted recently that our backscatter detector had } slipped and was almost touching the secondary detector though this has been } corrected and is no longer a problem. } Analysis of copper/gold standards are giving good quantitative results. } } I realize that these details are somewhat sketchy but does anyone recognize } this problem? Any input is greatly appreciated. } } Wayne England } ORTECH CORP. } Mississauga, Ontario } wengland-at-ortech.on.ca
Wayne,
Are you saying a) that the count rate changes as you adjust the SEM controls for brightness/contrast while doing an analysis at a stationary point or b) are you slowly scanning (or taking a series of multi-point analyses) over an area and seeing the count rate change as the brightness/contrast of the specimen image naturally changes?
If a), then you have a problem, but if b) then this is exactly what you would expect - either the composition (if a flat polished surface) and/or topography (if a rough surface) of the specimen is changing. Both will affect the EDX count rate.
Any Sr giving rise to a L alpha peak that might confuse with Si would also produce a much stronger K alpha/beta pair - as they don't show, you don't have any Sr. Similarly M lines from heavier elements would also produce K lines up around 8 keV, so I doubt that you have any other element genuinely contributing to the spectra.
What count rates are you running at? If you have a high count rate, then an escape peak from an element around 3.5keV might screw up the Si K peak, for example, lots of Sb, Sn or Ca but as you don't mention these, I presume this is not an issue.
You might have also have some real problems:
1. You may have a (maybe more than one) ground loop. Check that all your ancilliary equipment is getting its power from the same source. Preferably, this means connecting eveything to the auxillary power outputs of your SEM. Also make sure that you don't have any spurious ground links - for example, when the EDX detector is fully disconnected from the analyser, it should be isolated from the SEM. If the detector is grounded both directly through the SEM and via its own analyser, you will get odd things happening.
2. You say everthing is fine for Cu and Au standards. Are you using the Au M and Cu L? You need to check using a reference with peaks in the same part of the spectrum as the region you are having problems in since the problems you are experiencing may not be simple, by which I mean, the effect may have a complex relationship to energy, mainly affecting only the low engery region of the spectrum. However, if these standards are genuine, nice flat polished standards and the situation is as b) above, then this is exactly what you would expect.
Hope that helps. If it raises more questions, come back to me.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
OK, I'll jump in the middle of this. We have a digital camera also. We have one of the referenced Film Scanners as well as three 35mm film scanners. We also have flatbed scanners.
The bit depth,cost,dpi optical resolution, versatility and ease of use of any of our scanners and others on the market outweigh any scan line issues as well as the expense of getting the equivalent resolution and bit depth of scanners.
John D. Warren Southern Sales Manager "see what develops" Digital Photographic Imaging Group Polaroid Corporation
4525 Leonard Parkway Richmond, Virginia 23221-1809 Office 804.254.1011 Fax 804.254.1013 Internet warrenj1-at-polaroid.com
At 12:40 15/10/96 -0600, you wrote: } } I am considering purchasing a negative film scanner and I have narrowed the } choices to the Polaroid Sprintscan 45, the Nikon LS4500AF or the LeafScan } 45. The LeafScan 45 is no longer produced but I may be able to get a hold of } one of the last ones or a rebuilt one. I have used the LeafScan 45, but I } have no experience with either the Polaroid or the Nikon. The LeafScan } exceeds the performance of the other two (according to the specifications), } but is the cost justified? The Polaroid and Nikon are approximately half the } price of the LeafScan. } } I would appreciate hearing your comments (advantages/disadvantages) if you } have used either the Polaroid or the Nikon, especially if you have } experience with the LeafScan for comparison purposes. } Thanks in advance. } ******************************************************* } David F. Teter } Los Alamos National Laboratory } Materials Science and Technology: Metallurgy (MST-6) } Mail Stop: G755 } Los Alamos, NM 87545 } ph: (505)665-0160 fax: (505) 665-0657 } e-mail: teter-at-lanl.gov } ******************************************************* David & all: I have an interest in digital cameras - we sell the MicroLumina in Australasia. However, I truly believe that a high resolution digital camera, although more expensive, is a better solution than a scanner. I don't like scanners, I've had too much trouble with scan lines etc. Combined with a macro lens a digital camera can copy negs on a light box beautifully, it can also "copy" prints, apparatus, people and record microscope images. In short it's infinitely more versatile. Jim Darley
Probing & Structure } (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } A great microscopy site http://www.ultra.net.au/~pns/
I would agree with Geoff Avery that the Robinson detector will give you better atomic number contrast than the four quadrant detectors but don't discount the latter totally. We use a Kevex 4QBSE and have had extremely good results from this. (in previous employment I used a Robinson setector). The 4QBSE detector certainly achieves the manufacturers specs and has given no problems in the time we have had it (6 years). When I used the Robinson type detector an occassional anoyance was that on occassions I had to remove the detector from the SEM in order to achieve high res SE images on certain larger specimens.
I would say that a lot depends on what else you have and want. For example we have a retractable CL detector and so could not use a Robinson without physically swapping the detectors in the same port. (not a good idea)
Hope this helps
Pete
} Dear Microscopists, } } I was wondering about the kinds of BSE detector systems that people have } with their SEM's. What are your opinions? How reliable is the detector } system that you have? Which one/one's would you recommend? } } Thanks in advance, } Peling Melville } } -------------------------------------------------------------- } Peling Fong Melville } Senior Scientific Assistant } Interdepartmental Facilities } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024-5192 U.S.A. } ****************************** } E-mail: peling-at-amnh.org } Work #: (212) 769-5469 } FAX #: (212) 769-5495
************************************** * Pete Ainsworth * * Dept. Geology & Applied Geology * * Lillybank Gardens * * University of Glasgow * * Glasgow G12 8QQ * * e-mail: ainswort-at-geology.gla.ac.uk * * Tel : 0141 330 5505 (direct) * **************************************
} Dear all, } } A colleague would like to study the development of polysaccharide biofilms } (secreted by Pseudomonad bacteria) on lettuce leaves. She asked whether } there were any quick and easy (or, if necessary, long and tricky) ways of } seeing whether these polysaccharides were present on leaves before and } after they are turned into salads. My suggestion was to use a simple } polysaccharide stain on hand sections of leaves, but the film may be too } thin to distinguish it from the cuticle, even if she compared leaves from } sterile cultures with field-grown leaves. Any suggestions? } } TIA, } } Rosemary White } Department of Ecology and Evolutionary Biology } Monash University, Clayton, Victoria 3168, Australia } phone 61-3-9905 5670 email r.g.white-at-sci.monash.edu.au } fax 61-3-9905 5613 or rgwhite-at-vaxc.cc.monash.edu.au } } Rosemary, in the paper cited below, the author was able to distinguish between polysaccharides secreted by fungi and bacteria from those secreted by the plant's root tissues. This paper could be a good starting point. Citation: R.C.Foster. The Ultrastructure and Histochemistry of the Rhizosphere. New Phytol. (1981) vol. 89, 263-273. I hope this helps. Hank Adams EML New Mexico State Univ.
We, too have a Robinson-type detector, on our field emitter and get excellent results, especially with pesky samples that have local charging: multi-phase glasses and ceramics, loose rare-earth phosphor particles, silicone deposits on metals, etc. It is invaluable in these cases.
The only "problems" we have are: Scratching of the aluminum over coat by samples (operator error, not instrument problem) The necessity to retract the rod when it is necessary to get closer than 12 mm to the pole piece (it has to go someplace to collect electrons) "Buck-shot" noise on the CRT when the contrast is very high (just turn it down) No opportunity to select/add/subtract, etc. individual channels as in a plate (nature of the beast)
------------------------------------------------- Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-rd.sylvania.com
Our web sites: www.sylvania.com www.osram.de www.siemens.com
Dear Microscopists,
I was wondering about the kinds of BSE detector systems that people have with their SEM's. What are your opinions? How reliable is the detector system that you have? Which one/one's would you recommend?
Thanks in advance, Peling Melville
-------------------------------------------------------------- Peling Fong Melville Senior Scientific Assistant Interdepartmental Facilities American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 U.S.A. ****************************** E-mail: peling-at-amnh.org Work #: (212) 769-5469 FAX #: (212) 769-5495
} The Minnesota Microscopy Society is involved in a preliminary trial of } the Project Micro Program. One of the experiments calls for examining } various sands at low magnification (10x-20x). These sands should have } unique morphological or color properties which could be seen at this } magnification. If anyone has access to some unique sand we would } appreciate the help. Please contact me at my EMail address: } } RGRappe-at-mmm.com
} Would you be interested in sand collected by my well pump filter? The } grains are sharp edged and irregular - about 100 microns long. The } sand is predominantly an even mix of quartz (clear/white) and nearly } black particles. I forget the composition of the black - I think it } was comprised of O, Si, Fe, and maybe(?): Mg, Ca, Mn, and or Ti?? } A geologist friend belives it comes from decomposing (poorly } "sintered") igneous(sp?) rock which was once an undersea deposit of } volcanic ash.
I hope that everyone agrees that sand samples for MSA's middle school educational outreach program are an appropriate use of the listserver. I think that it's a great idea, and I'll be glad to receive labeled samples, for future distribution. Please send samples first to Rappe - he needs them now.
Caroline Schooley Educational Outreach Coordinator, Microscopy Society of America Box 117 (45301 Caspar Point Road) Caspar, CA 95420 Phone/FAX: (707)964-9460 Email: schooley-at-mcn.org Web: http://www.MSA.microscopy.com/ProjectMICRO/Books.html
A Philips EM301 is available for immediate delivery.
The instrument has been on a service contract and has been upgraded with a goniometer stage and diffusion pump.
For more information, please contact:
************************************************************************* Lucille A. Giannuzzi, Ph.D. Assistant Professor
Dept. of Mechanical, Materials, and Aerospace Eng.
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA *************************************************************************
I would like to have contact with peolple who are geographically close to New Orleans who have experience in the installation and configuration of imaging products such as frame grabber boards, computers, software & CCD cameras, for light microscopy applications.
_______________________________________________ SCOTT SCIENTIFIC P.O. Box 66552 Station Cavendish Montreal, Quebec, H4W 3J6, Canada
A colleague wishes to produce sphaeroplasts (wall-less bacteria) using lysozyme on cultures of E. coli. Is there a stain capable of differentiating between normal, walled E. coli cells and sphaeroplasts? With gram positive cells this is possible with a simple gram stain but this won't work with E. coli since it is already gram negative. Thank you.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[[ POSITION OPEN ]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]]
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South Bay Technology, Inc. a small and rapidly expanding laboratory equipment manufacturer is seeking an innovative, organized and self-motivated individual for a position in technical sales.
Responsibilities The successful candidate will report directly to the Vice President of Sales and Marketing and will be responsible for telephone follow up and qualification of prospects from leads generated through trade shows, journal advertising, industry trade groups, etc. The successful candidate will also make regular contact with existing customers informing them of new product developments and technical information specific to their application. The position will require extensive telephone contact and interaction with customers. The position will also involve responding to quotation requests, taking incoming calls for information and intitial processing of orders.
Qualifications Strong interpersonal, organizational and writing skills are absolutely required as is the ability to work as a member of a team. The candidate should have strong computer skills -familiarity with ACT!, Microscoft Word, Microsoft Excel, Microsoft Powerpoint and other software packages is desirable. Scientific background and/or familiarity with mechanical drawings is a plus.
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Greetings Folks, I have a question regarding Transmission Electron Microscope Film. A client of mine came up with several old boxes of DuPont Cromolar Electron Microscope Film. Type EM-7. It was outdated but had been stored in a freezer. We can find no information as to the speed of the film or other characteristics. I suppose it can be developed in D-19, like Kodak film but don't know for sure. Any information about this stuff would be greatly appreciated. Thanks much for your time.
Alex Greene Scientific Instrumentation Services, Inc. Austin, Texas
We have the Oxford Instruments TETRA BSe collector and it is fine down to an accelerarting voltage of ca. 4-5kV. Philips do a "low voltage BSE collector but I have no idea how well it works. Has any one out there got or know of a BSE collector which works ar a 1-2 kV or are we up against the dreaded Laws of Physiscs.
Patrick Echlin Multi-Imaging Centre CambridgeOn Thu, 17 Oct 1996, Peling Melville - Interdepartmental Facilities wrote:
} Dear Microscopists, } } I was wondering about the kinds of BSE detector systems that people have } with their SEM's. What are your opinions? How reliable is the detector } system that you have? Which one/one's would you recommend? } } Thanks in advance, } Peling Melville } } -------------------------------------------------------------- } Peling Fong Melville } Senior Scientific Assistant } Interdepartmental Facilities } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024-5192 U.S.A. } ****************************** } E-mail: peling-at-amnh.org } Work #: (212) 769-5469 } FAX #: (212) 769-5495 } } }
I am soon going to start working in a project that involves autoradiography on the electron microscopical level. There is not very much written about it (I've read Baker), so I have quite a few questions... One of my questions is regarding exposure time; I'll be working with a gamma-radiator (In-111).
If you have any idea, please contact me!
My regards,
Margareta Halin Dept. of anatomy and histology SLU Sweden
Message-Id: {9610211749.AA3209-at-czjnotes03} To: spm {spm-at-di.com}
Fellow microscopists,
Carl Zeiss plans the launch of a Scanning Optical Near Field Microscope (SNOM) in early 1997.
Everybody who is interested in this development may leave his address on our web-page http://www.zeiss.de/mi/rsm/form.html and will get further informations as soon as they are released.
Sincerly,
Stefan Mueller-Pfeiffer
------- Stefan Mller-Pfeiffer, Carl Zeiss Jena GmbH, Microscope Division
Actually, there's a considerable amount of older information on the technique itself. M. Salpeter contributed the most to this very tedious and laborious technique, and has a very good book on the subject. She also has several chapters in some of the EM techniques books, such as ADVANCED TECHIQUES IN BIOLOGICAL ELECTRON MICROSCOPY (J.K. Koehler, ed.).
This should get you started if you REALLY want to do this.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia
I have a feeling that what you are looking for is the new CENTARUS BS detector made by Mike Cowham of KE Developments in Toft. Although I have not yet been able use one I do understand that it operates well down to about 600 V - which is good and so is ideal for LV SEM. It also has a number of other advantages over the Robinson type. One problem I have seen with the Robinson is having to replace the scintillator after a number of years. With heat from the beam one finds the scintillator develops cracks which affects efficiency. A costly exercise.
Mike can be e-mailed at CompuServe: 100610.2353
I look forward to attending your Cryo workshop.
Best regards
Dane Gerneke E M Unit University of Cape Town Tel +27 21 650 2819 Fax +27 21 689 1528 dane-at-uctvms.uct.ac.za
University of Oxford on 7-10 April, 1997 ----Abstract deadline: 2 December 1996
Organized on behalf of the Royal Microscopical Society by: **Prof Anthony G Cullis (a.g.cullis-at-sheffield.ac.uk) **Dr John L Hutchison (john.hutchison-at-materials.ox.ac.uk)
Co-sponsored by the Institute of Physics (EMAG) and Endorsed by the Materials Research Society
CENTENARY OF ELECTRON DISCOVERY ------------------------------------------------------------
The conference will feature a special Symposium to celebrate the centenary of the discovery of the electron. Key presentations will be given by: Dr W F Brinkman (Vice-President for Physical Sciences Research, Bell Labs, Murray Hill) **The Materials Behind the Telecommunications Revolution Dr T Matsuo (Managing Director of Semiconductor Equipment Division, JEOL Ltd, Tokyo) **The Evolution of Semiconductor E-Beam Lithography and Metrology Prof K van der Mast (Philips Electron Optics, Eindhoven) **The Development of Electron-Optical Imaging and Diffraction Systems
MAIN CONFERENCE SCIENTIFIC SESSIONS
These will focus on the most recent advances in the application of transmission and scanning electron microscopy, X-ray diffraction and SPM to the study of the structural and electronic properties of semiconducting materials.
Main topic areas: - Characterisation of as-grown semiconductors. - Investigation of lattice defect and impurity behaviour. - Study of the effects of semiconductor processing treatments. - Assessment of finished electronic devices.
Special conference sessions: - Developments in high resolution transmission electron microscopy. - The nature of epitaxial layers, including quantum well, wire and dot st= ructures ---- strain relaxation, defect introduction, morphological distortion, self-organization, luminescence. - The structure and properties of dislocations and grain boundaries. - Metal-semiconductor contacts and silicides. - The effects of processing treatments. - The exploitation of advanced scanning techniques ---- SEM-EBIC, SEM-CL, etc ---- STM, AFM, BEEM, etc.
INVITED SPEAKERS -----------------------------
Prof P J Goodhew (University of Liverpool) **Dislocation Behaviour in Strained Layer Interfaces Prof R J Hamers (University of Wisconsin-Madison) **STM Studies of CVD Processes on Si Surfaces Dr D C Houghton (Canadian National Research Centre, Ottawa) **Advances in Epitaxial Strained Layer Devices Dr D E Jesson (Oak Ridge National Laboratory, Tennessee) **Exploring Instabilities and Metastabilities in Semiconductor Growth Dr J-L Rouvi=E8re (CEN, Grenoble) **GaN Growth: Influence of Polarity and Strain Prof J C H Spence (Arizona State University) **Dislocation Kink Behaviour in Semiconductors Prof H P Strunk (University of Erlangen) **Self-Organization and Defect Mechanisms in Heteroepitaxial Growth Prof S Takeda (University of Osaka) **The Structures of Extended Defects in Si and Ge Analysed by HRTEM Dr R T Tung (Bell Laboratories, Murray Hill) **Control of Silicide Layers in ULSI Devices: Simple Principles at Work Dr J Vanhellemont (IMEC, Leuven) **TEM Studies of Processed Si Device Materials Dr P R Wilshaw (University of Oxford) **Developments in SEM:EBIC Studies of Semiconducting Materials
********************************
The Proceedings of the conference will be published. Further details, including registration and abstract submission information, can be obtained from: The Administrator, The Royal Microscopical Society, 37/38 St Clements, Oxford OX4 1AJ, UK Tel: +44-(0)1865-248768 Fax: +44-(0)1865-791237 E-mail: meetings-at-rms.org.uk WWW: http://www.rms.org.uk
Message-Id: {199610211727.KAA10516-at-goodguy.goodnet.com} Comments: Authenticated sender is {earosen-at-mail.goodnet.com}
I just have one question to ask you all what the job market is lately for a biologist with a M.A. or M.S., or B.S. degree?? I am doing some research into what major I should choose for school and really like biology but have hard mixed reviews about the job market...
Thanks in Advance.....
\\|// (o o) ~~~~~~~~~~~~oOOo~(_)~oOOo~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
{ { {This message is made of 100% recycled electrons} } }
Cheers ;o) :o) %o) Eric http://www.indirect.com/www/earosen/index.htm
We have an YAG crystal BSE detector modified by Dr. Autrata, Czech Republic. The BSE detector is installed on a Hitachi S-900 LVSEM and is capable of detecting 1nm immuno-gold signal. The efficient voltage can be down to 2.6kV.
What technology is used? Is saturation a problem when using high potentials and currents? Woody ______________________________ Reply Separator _________________________________
I have a feeling that what you are looking for is the new CENTARUS BS detector made by Mike Cowham of KE Developments in Toft. ...snip... Mike can be e-mailed at CompuServe: 100610.2353
I look forward to attending your Cryo workshop.
Best regards
Dane Gerneke E M Unit University of Cape Town Tel +27 21 650 2819 Fax +27 21 689 1528 dane-at-uctvms.uct.ac.za
I've got a surfeit of Polaroid 545 (4"X5") film backs and am looking to trade one or two for the same size back, but pack film format. I shoot a lot of Polaroid 553 and could really use another one. Anyone want to do an even exchange? Please reply to DRStad-at-Juno.com.
I gave a seminar last Friday on high resolution SEM of chloroplast ultrastructure. At the conclusion of my talk I got a question that I have gotten before: "How do you know that what you are seeing is not an artifact of fixation?" The answer I always use is: 1) fixation artifacts are rather apparent so if there was one (membrane blebbing, broken membranes, etc) I would have discarded the samples. 2) We have been looking at chloroplasts for over 40 years under a wide variety of fixation conditions and if the structure as we see it is an artifact, it is an amazingly consistent one. 3) The structural data are consistent with biophysical, bioenergetic and biochemical data taken on similar systems.
But this got me thinking so I ask the following question to you all...
Does anyone have an example of an artifact in either SEM or TEM that seriously mislead the scientific community? I don't wish to get into criticizing other people's work, but in the 41 years since the invention of the ultramicrotome (the real start of biological TEM) and the 30 years since the commercial availability of the SEM, how much impact on scientific understanding have artifacts had or how much impedance have they provided to science? What about those of you doing high pressure freezing? What are the differences you see in samples prepared by HPF vs. standard chemical fixation?
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
} Does anyone have an example of an artifact in either SEM or TEM that } seriously mislead the scientific community? I don't wish to get into } criticizing other people's work, but in the 41 years since the invention of } the ultramicrotome (the real start of biological TEM) and the 30 years } since the commercial availability of the SEM, how much impact on scientific } understanding have artifacts had or how much impedance have they provided } to science? What about those of you doing high pressure freezing? What } are the differences you see in samples prepared by HPF vs. standard } chemical fixation?
I'm not sure about "seriously" misled the community, but....
First example - Geoff Hyde showed that spore formation in the fungus Phytophthora was by membrane extension around nuclei, not by fusion of lined-up membrane vesicles. He saw this with rapid plunge freezing (I think), not with high pressure freezing, nor with conventional chemical fixation. People had scratched heads for a long time as to how these membrane vesicles managed to line up so nicely, and here, finally was the answer - they didn't! THe next question is - what guides the membranes? The answer is probably some component of the cytoskeleton - I think he showed that things are messed up with cytochalasin or oryzalin.
Second example - John Pettit looked at mouse kidney tissue (?organ? - this is a botanist speaking) in which it was thought that salt secretion was via vesicles (seen after chemical fixation) that were all lined up, and somehow they fused with the membrane at a particular spot when secretion was induced. After slam freezing, instead of lined-up vesicles, in serial sections he saw long curly membranous tubes shaped a bit like the cord that connects your telephone ear/mouthpiece to the body of the phone. Active salt secretion was associated with slight untwisting of the tubes together with a slight increase in their diameter. Fascinating.
I suspect there are quite a few chemical fixation artefacts like this that can be avoided with various freezing techniques - as long as you can get your tissue small enough to avoid ice damage, or can trim it down without wounding artefacts.....
cheers,
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Clayton, Victoria 3168, Australia phone 61-3-9905 5670 email r.g.white-at-sci.monash.edu.au fax 61-3-9905 5613 or rgwhite-at-vaxc.cc.monash.edu.au
Two positions are available at the University of Central Florida. The new Joint Lucent/UCF Materials Characterization Facility currently consists of a JEOL 2000FX TEM with EDS, Philips EM400T, specimen preparation facility, JEOL 6400 SEM, JEOL 733 electron microprobe, Riber SIMS, Tandentron RBS. A Phi Auger, Cameca SIMS, and a Hitachi S-800 SEM will be added within the year. The facility is located in nearby Research Park.
An electron microscope engineer is desired with an expertise in servicing and running electron microscopes and related equipment. Responsibilities will include servicing equipment, training students, and aiding faculty and staff with analytical procedures.
A Post-Doctoral/Research Associate is desired with an expertise in (analytical) transmission electron microscopy, and advanced specimen preparation of materials. Responsibilities will include performing independent research, training students, and assisting with graduate student research projects.
UCF is located near Lucent Technologies (formely AT&T), Westinghouse, Lockheed Martin, NASA Kennedy Space Center, Walt Disney World, Universal Studies, Harris, and others.
Candidates for either position should submit a resume (please specify the position your are applying for) and at least three references, to Dr. Lucille A. Giannuzzi, Mechanical, Materials & Aerospace Engineering, University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450.
UCF is an affirmative action/equal opportunity employer. As an agency of the State of Florida, UCF makes all materials available for public review.
************************************************************************* Lucille A. Giannuzzi, Ph.D. Assistant Professor
Dept. of Mechanical, Materials, and Aerospace Eng.
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA *************************************************************************
Green Box?? If so, I have used this film a long time ago. The emulsion settings are very similar to 4489 film. Try a few different settings. You can develope in same D-19 solutions as 4489. I was very mad when this film was no longer avaiable, it was about $20/box cheaper than kodak film. Oh well.
Best of luck, Ed Calomeni Medical College of Ohio Dept. Pathology Toledo, OH 43699 emlab-at-opus.mco.edu
I work with crustacean ultrastructure. In a number of species we have noted structures that look just like the multi-lamellar bodies associated with surfactant in mammalian lung. Although I (along with colleagues at other institutions) am convinced that these are not fixation artifacts, but rather represent real structures, there are many who do believe that we have fixation artifacts (with justification).
How could we ever prove otherwise?
Additional responses would be welcomed.
Don
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey * fax: (609) 771-2674 Trenton, NJ 08650-4700
(* formerly Trenton State College; please note our new name)
I am working with a diffusible tracer, C14-iodoantipyrine and need to section the tissue. After the experiment the head is frozen in methylbutane at dry ice temp, freeze dried and then the area of interest is dissected out. This leave a specimen of } 1 mm sq. I need to then section the tissue at 20 um. I have seen some older references where the tissue is placed directly into degassed molten paraffin. Has anyone had any experience with this technique and willing to pass along their experiences?
} Does anyone have an example of an artifact in either SEM or TEM that } seriously mislead the scientific community? } } Bob
This may still be a matter of controversy, but off the top of my head, I would suggest mesosomes in bacteria. Many people were excited about them when first seen, but they have come to be regarded as fixation artifacts, especially from glutaraldehyde. In SEM, I would regard any measurement made on soft tissue as artifactual, due to both fixation/dehydration/drying and the physics of how SEMs work. I'm leery of any measurements, including TEM or LM, of processed tissues generally. Phil
&&&&&&&&&&& Illigitmi non carborundum &&&&&&&&&&&
Philip Oshel *all mail:* Microscopy University of Illinois Station A Rm 74 Bevier Hall PO Box 5037 905 S. Goodwin Ave. Champaign, IL 61825-5037 USA Urbana, IL 61801 (217) 244-3145 oshel-at-ux1.cso.uiuc.edu (217) 355-1143 home
In my memory, the most widespread instance of a (purported) fixation artifact misleading a large proportion of the scientific community was that of the proposed microtrabecular system which was supposed to be common to all eukaryotic cells. Observations of thick sections of glutaraldehyde-fixed cultured cells by HV-TEM revealed a very fine wispy network suspended in the cytosol and numerous functions (mostly intracellular transport and communication) were attributed to it. Many imminent scientists were involved with it and many careers were made studying it. Its validity was always questioned by another side which (I think) ultimately demonstrated that it was inducible by chemical fixation. Anyway, cell biology books of 10-12 years ago covered it, and its absent from them today. I can probably direct you to specific literature if you are that interested.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia
Posted-Date: Tue, 22 Oct 1996 14:39:42 -0400 (EDT) Message-ID: {01BBC026.3DB84F80-at-OAKMONT.SEAS.UPENN.EDU}
Hi
Does anyone know how to reach a company called Microporous Materials = Limited in Braunston, UK, preferrably a fax number or email address? = They make a polystyrene bead with porous structure. (I am trying to = find or develop beads that have a density less than water. No one seems = to have anything less than 1.02 g/cm^3.)
Thanks
Eric Johnston Department of Bioengineering University of Pennsylvania P: 215-898-1958 F: 215-573-2071 ericdj-at-eniac.seas.upenn.edu
Eric, What kind of job you get depends on what field of biology you want to study and what part of the country you are willing to work in AND what kind of salary you are looking for. There are a fairly good number of jobs in the medical support field, for instance, but you need to be where there is a large medical center and you can't be looking for big pay... I am in a graduate school where many students graduate with a M.S. in speech pathology or audiology. Here, the field of speech is just opening up, so there are a fair number of jobs in the area. But this local area is saturated with audiologists, so to get a good job, graduates have to be willing to relocate. Also, the audiologists are talking about switching to a 4 year, post-graduate degree for certification in the future, so by the time you got to gradute school, things could be different. The best way, I think, to approach this is to decide what you would enjoy doing and how much money you want to start your carrier making. Don't limit your choices by how long you would be in school. I did that and here I am, 15 years later, getting the degree to do what I wanted way back when...
Good luck, Karen Pawlowski
On Mon, 21 Oct 1996, Eric wrote:
} I just have one question to ask you all what the job market is lately } for a biologist with a M.A. or M.S., or B.S. degree?? I am doing } some research into what major I should choose for school and really } like biology but have hard mixed reviews about the job market... } } Thanks in Advance..... } } } \\|// } (o o) } ~~~~~~~~~~~~oOOo~(_)~oOOo~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } { { {This message is made of 100% recycled electrons} } } } } Cheers ;o) :o) %o) } Eric } http://www.indirect.com/www/earosen/index.htm }
In message {199610190251.VAA02487-at-tristero.io.com} "A. Greene" writes: } Greetings Folks, I have a question regarding Transmission Electron } Microscope Film. A client of mine came up with several old boxes of DuPont } Cromolar Electron Microscope Film. Type EM-7. It was outdated but had been } stored in a freezer. We can find no information as to the speed of the film } or other characteristics. I suppose it can be developed in D-19, like Kodak } film but don't know for sure. Any information about this stuff would be } greatly appreciated. Thanks much for your time. } } Alex Greene } Scientific Instrumentation Services, Inc. } Austin, Texas } } [INDEPENDENT ELECTRON MICROSCOPE MAINTENANCE] } Hi Alex,
I used to use that film. I developed it in D-19, full strength, for 3.0 minutes, stop bath 30 sec, Kodak Rapid Fixer for 3.0 minutes, hypoclear 1.0 min, 10 minute minimum water wash. HOWEVER, the 3.0 minute development time only works if you calibrate your TEM exposure so that you get a good negative at 3.0 minutes. Your TEM must have a way to set a "film speed" parameter, like my Philips CM12 TEM does. On my former Philips EM300, there was a film sensitivity control. Most TEM's have a way of measuring electron brightness which is metered somehow and that meter output can be adjusted by the film sensitivity control.
So if you want to standardize to 3.0 minute development time, then you need to do a few exposure tests to get a good full toned negative at 3.0 minutes.
Alternatively, if you want to dilute D-19 to 1:1 or 1:2, then perhaps standardize on 4.0 or 5.0 minute development and calibrate your TEM exposure accordingly.
Hope this helps!
Gib
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 Its over, but not forgotten, and it was a blast!
In reply to the query about chloroplast ultrastructure in the SEM, you need to distinguish between artefact and damage. I use the term artefact to describe an altered state ie some which is there in a natural state but s "seen" in an altered state. In that respect all EM is an artefact. BUT what we try to do is to understand the nature of the transformation from the wet organic reality to the dry inorganic material from which the image is derived.Sample preparation should be considered as constructive fossilization. Damage is where we the operaters muck things up.So the bottom line is: Artefact is inevitable and needs careful understanding Damage is inexcsible.
Hope this helps
Patrick Echlin Cambridge UKOn Mon, 21 Oct 1996 wise-at-vaxa.cis.uwosh.edu wrote:
} To all, } } I gave a seminar last Friday on high resolution SEM of chloroplast } ultrastructure. At the conclusion of my talk I got a question that I have } gotten before: "How do you know that what you are seeing is not an } artifact of fixation?" The answer I always use is: 1) fixation artifacts } are rather apparent so if there was one (membrane blebbing, broken } membranes, etc) I would have discarded the samples. 2) We have been } looking at chloroplasts for over 40 years under a wide variety of fixation } conditions and if the structure as we see it is an artifact, it is an } amazingly consistent one. 3) The structural data are consistent with } biophysical, bioenergetic and biochemical data taken on similar systems. } } But this got me thinking so I ask the following question to you all... } } Does anyone have an example of an artifact in either SEM or TEM that } seriously mislead the scientific community? I don't wish to get into } criticizing other people's work, but in the 41 years since the invention of } the ultramicrotome (the real start of biological TEM) and the 30 years } since the commercial availability of the SEM, how much impact on scientific } understanding have artifacts had or how much impedance have they provided } to science? What about those of you doing high pressure freezing? What } are the differences you see in samples prepared by HPF vs. standard } chemical fixation? } } Bob } } } Robert R. Wise, PhD } Director, UWO Electron Microscope Facility } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-uwosh.edu } } }
I am a student in the Northwestern business school (Kellogg). We are doing some research for the McCrone Research Institute. Our goal is to find who could use the following item and what would be a reasonable price. They have a particle atlas on CD-ROM (2,000 particle images), with a glossary of terms and techniques.
Any, and all, comments would be very much appreciated. Questions and comments should be directed to me at:
You are pretty selective on what you call an artifact (blebbing, broken membrane). One only has to compare an aldehyde fixed mitochondrion with a quick frozen one. In quick frozen tissues, mitos tend to be smaller, much more condensed. somebody showed that if you pre-treat the tissue with an uncoupler of ox-phos, the quick frozen ones look like aldehyde fixed tissue. In my own work with cholinergic nerve terminals in torpedine rays, we showed that aldehyde fixed nerve terminals had no vesicles along the pre-synaptic membrane but quick frozen tissues had a zillion. Aldehyde fixation caused calcium entry and loss of active zone vesicles. Van Harreveld showed the classic view of aldehyde fixed brain with no spaces between anything is an artifact of fixation; quick frozen brain shows much more extracellular space. Microtubules are lost if one fixes at 4 C but not if you fix at room temp. Nuclear volume varies significantly depending on the fix and dehydration protocol. There are lots of other examples. It is extremely dangerous to assume what one sees (and even prefers visually) after any type of fixation is totally unaltered.
} To all, } } I gave a seminar last Friday on high resolution SEM of chloroplast } ultrastructure. At the conclusion of my talk I got a question that I have } gotten before: "How do you know that what you are seeing is not an } artifact of fixation?" The answer I always use is: 1) fixation artifacts } are rather apparent so if there was one (membrane blebbing, broken } membranes, etc) I would have discarded the samples. 2) We have been } looking at chloroplasts for over 40 years under a wide variety of fixation } conditions and if the structure as we see it is an artifact, it is an } amazingly consistent one. 3) The structural data are consistent with } biophysical, bioenergetic and biochemical data taken on similar systems. } } But this got me thinking so I ask the following question to you all... } } Does anyone have an example of an artifact in either SEM or TEM that } seriously mislead the scientific community? I don't wish to get into } criticizing other people's work, but in the 41 years since the invention of } the ultramicrotome (the real start of biological TEM) and the 30 years } since the commercial availability of the SEM, how much impact on scientific } understanding have artifacts had or how much impedance have they provided } to science? What about those of you doing high pressure freezing? What } are the differences you see in samples prepared by HPF vs. standard } chemical fixation? } } Bob } } } Robert R. Wise, PhD } Director, UWO Electron Microscope Facility } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-uwosh.edu
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Lots of times this has happened but my life is a bit too busy to go into much detail. Look at the mesosomes in bacteria, there were actually fixation artifact, found when free fracture was done. The pore complex in basidiomycetes and ascomycetes in fungi. and there are plenty more. The cell membrane in bacteria often is artifactual shown true by free fracture where there is no fixation.
Good luck, its actually fun to make a list and look through the literature for them as I have my students do.
Judy M.
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
Phone: 209/474-5284 FAX: 209/474-5600 e-mail: murphy-at-sjdccd.cc.ca.us program web page: http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html
I gave a seminar last Friday on high resolution SEM of chloroplast ultrastructure. At the conclusion of my talk I got a question that I have gotten before: "How do you know that what you are seeing is not an artifact of fixation?" The answer I always use is: 1) fixation artifacts are rather apparent so if there was one (membrane blebbing, broken membranes, etc) I would have discarded the samples. 2) We have been looking at chloroplasts for over 40 years under a wide variety of fixation conditions and if the structure as we see it is an artifact, it is an amazingly consistent one. 3) The structural data are consistent with biophysical, bioenergetic and biochemical data taken on similar systems.
But this got me thinking so I ask the following question to you all...
Does anyone have an example of an artifact in either SEM or TEM that seriously mislead the scientific community? I don't wish to get into criticizing other people's work, but in the 41 years since the invention of the ultramicrotome (the real start of biological TEM) and the 30 years since the commercial availability of the SEM, how much impact on scientific understanding have artifacts had or how much impedance have they provided to science? What about those of you doing high pressure freezing? What are the differences you see in samples prepared by HPF vs. standard chemical fixation?
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
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At 10:54 22/10/96 -0400, you wrote: } } I work with crustacean ultrastructure. In a number of species } we have noted structures that look just like the multi-lamellar bodies } associated with surfactant in mammalian lung. Although I (along with } colleagues at other institutions) am convinced that these are not } fixation artifacts, but rather represent real structures, there are many } who do believe that we have fixation artifacts (with justification). } } How could we ever prove otherwise? } } Additional responses would be welcomed. } } Don } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } The College of New Jersey * fax: (609) 771-2674 } Trenton, NJ 08650-4700 } } (* formerly Trenton State College; please note our new name) ************************************************ Don have you tried staining GA only fixed tissue with Sudan Black B or another lipid stain? Over 30 years ago it was shown that poorly fixed lipid droplets can form myelin-like, concentric artificial membranes. Alternatively you could treat the sections with lipase and digest any lipids. I rather expect that you are seeing such lipid droplets. However, if those structures are "real", chances are that they are lipid based anyway. Regards Jim Darley
Probing & Structure } (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } A great microscopy site http://www.ultra.net.au/~pns/
I currently use it, and find it worth every bit worth the price it was.
Hope that helps.
Shalom from Jerusalem, Azriel Gorski
+++++++++++++++++++++++++++++++++++++++++++++++ Azriel Gorski, Head Optical Microscopy Laboratory Division of Identification and Forensic Science Israel National Police 91906 Jerusalem
On Tue, 22 Oct 1996, Brad Kort wrote:
} I am a student in the Northwestern business school (Kellogg). We are } doing some research for the McCrone Research Institute. Our goal is to } find who could use the following item and what would be a reasonable } price. They have a particle atlas on CD-ROM (2,000 particle images), } with a glossary of terms and techniques. } } Any, and all, comments would be very much appreciated. Questions and } comments should be directed to me at: } } 312/422-2293 (day) } 708/361-7613 (evening) } bkort1-at-xnet.com (anytime) } } Thanks in advance, } Brad Kort }
Dear Fellow Sufferers, I am trying to analyse sputtered films by TEM/EDXA. These contain a significant amount of Ar, but I don't have a suitable standard (isn't that strange!). I can calculate a theoretical k factor which will give a reasonable estimate, but that still leaves me scratching around for a decent peak profile. Has anyone any solutions to this problem? Regards, Eric
Dr Eric Lachowski University of Aberdeen Department of Chemistry tel +44 1224 272934 fax +44 1224 272921 e.lachowski-at-abdn.ac.uk
Message-Id: {9610230822.AA28654-at-sun1.biomed.cas.cz} Comments: Authenticated sender is {benada-at-sun1.biomed.cas.cz}
Dear colleagues, I should like to use Alcian blue for fixation of envelope of bacteria (A procedure which was reviewed by T.A. Fassel et al., 1992). I have problems with dissolving this dye. Although I have purified it according to M.A. Hayat (1989), I can'n prepared 0.5% stock solution without any precipitate. I an using Alcian blue 8 GS (8 GX) for Microscopy FLUKA, Cat.No.05500.
Could anyone give me some advice?
Thanking you in advance Your sincerely O. Benada +---------------------------------------------------------------+ Dr. Oldrich Benada Acad. Sci. CR Phone: +42-2-4752399 Institute of Microbiology Fax: +42-2-4715743 Electron Microscopy Group E-mail: benada-at-biomed.cas.cz Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +---------------------------------------------------------------+
Hi, everybody, I agree with Philip Oshel (message from 22.10.1996) that we cannot belive to the measurments of the fixed and especialy dehydrated tissues. Do somebody have an opinion about all cryo methods do they affect the size of cells and organels and how significant these changes are?
The current expanding discussion of examples of artifacts in EM hasn't mentioned that there's a book on the subject published in 1988 which could be usefully consulted called , not surprisingly,
Artifacts inBiological Electron Microscopy eds Richard Creng / Karen Klomparens Plenum, ISBN 0-306-42863-6
it isn't concerned too much with what's recently been alluded to ie. results from cryotechniques being probably the strongest evidence for determining what are artifacts in chemically prepared specimens - but it's worth a look for the uninitiated dealing with conventionally prepared specimens. It goes into some detail on a range of artifacts in TEM, SEM and allied techniques including those generated by improper use of the instruments themselves.
Laurence Tetley Dr Laurence Tetley IBLS EM Centre Joseph Black Building University of Glasgow Glasgow G12 8QQ
Some of you know more about the Particle Atlas CD than I do. Where is it available? Who does it? How much it costs? The last Particle Atlas I saw (and have) was published by Ann Arbor Science in 1973 (Second Edition). There should be something fresher out there... Thanks for any information I get.
************************************************************************* Dr. Kristof KOVACS Associate Professor University of Veszprem, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684 *************************************************************************
I am looking for price and availability information on thermal paper for Mitsubishi video copy processor model K65H. Please repond to me and not the entire net.
Bob
Robert R. Wise, PhD Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
Hi everybody, I need some advice. We would like to check for the proportion of vesicles which are outside out as opposed to inside out in liver plasma membranes. Does anyone know of, or has used, a cytochemical marking for TEM to selectively label: a) only the extracellular domain or b) only the intracellular domain of a typical common plasma membrane protein. In particular, has anyone used colloidal gold-tagged lectins to bind membrane glycoproteins? If so, do you have a protocol? Any advice will be appreciated including suggested readings. TIA Regards, John Gabrovsek CCF Cleveland, Ohio
We are currently upgrading our Image analysis software and are considering purchasing either Universal Imaging's Metamorph or COMPIX's C*Imaging product. And I was wondering wether any listserv subscribers have any experience with either product that might help us giude our decision. Thanks in advance Lloyd Williams Manager of Bio Imaging Facility Center for Study of Gene Structure and Function Hunter College e-mail williams-at-genectr.hunter.cuny.edu
The z-control in our Cambridge S240 refuses to budge. I was wondering if anyone had any helpful hints before we tackle it. Any help is greatly appreciated, since getting phone support has proven difficult. Thanks.
Jeff
------------------------------------------------------------------ -- Jeff Shield -- -- Assistant Professor -- -- Department of Materials Science and Engineering -- -- University of Utah -- -- Salt Lake City, Utah 84112 -- -- 801/581-3179 -- ------------------------------------------------------------------
I am a student in the Northwestern business school (Kellogg). We are doing some research for the McCrone Research Institute. Our goal is to find who could use the following item and what would be a reasonable price. They have a particle atlas on CD-ROM (2,000 particle images), with a glossary of terms and techniques.
Any, and all, comments would be very much appreciated. Questions and comments should be directed to me at:
X-Authentication-Warning: gw: Host [10.17.1.22] claimed to be Message-Id: {326D7DEB.66AA-at-inet.guthrie.org}
To Whom It May Concern: I am a recent graduate of Mansfield University (B.S. in Biology with a Chemistry minor) and a current student at Robert Packer Hospital's School of Medical Technology. At Mansfield Uni. I conducted undergraduate research on a malignant, murine macrophage cell line --- P388D1 (ATTC). I experimented with carbonyl iron uptake and cell cycle correlations. The only thing that I was unable to find with this particular cell line in my literature search was electron microscopy. If I had some background of its cellular and surface components, I would better understand the cell response from my research. At the current moment I intend to continue the research here at RPH in 1997 with a stronger emphasis on membrane receptors with the use of Flowcytometry. It would be greatly appreciated if I could find this information of electron microscopy of this cell line through your organization or through your reference to another. Thank you for your time.
Sincerely,
Tammy M. Walker
I can be contacted at: twalker-at-inet.guthrie.org
I believe that the particle atlas on CD-ROM would be very useful to people (like myself) in the medical device, pharmaceutical, and biotech fields. I routinely have to isolate and identify particle contaminants in and on such products, using various microscopic and spectroscopic techniques. The price should be comparable to (or hopefully less than) what we would pay for the book version.
-Bob ********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 USA ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com **********************************
I am a student in the Northwestern business school (Kellogg). We are doing some research for the McCrone Research Institute. Our goal is to find who could use the following item and what would be a reasonable price. They have a particle atlas on CD-ROM (2,000 particle images), with a glossary of terms and techniques.
Any, and all, comments would be very much appreciated. Questions and comments should be directed to me at:
Los Alamos will shortly be advertising for a technical staff member in the Electron Microscopy Facility. Here is the job description:
Summary:
Work in the Center for Materials Science, Materials Science and Technology Division at Los Alamos National Laboratory supporting a variety of basic energy sciences and other DOE programs. Provide expertise in electron microscopy and be part of a team that operates the CMS Electron Microscopy Facility. In particular take the lead in operating and maintaining a VG HB601UX field-emission transmission elecron microscope, a JEOL 3000F field-emission high resolution transmission electron microscope and a Philips CM30 analytical transmission electron microscope, along with associated parallel electron energy loss systems, x-ray energy dispersive systems and ccd cameras. Be involved in all aspects of thin film sample preparation. Teach and assist other employees and visitors in the operation of the microscopes. Be involved in research that uses the electron microscopes.
Required Skills, Knowledge, Abilities:
The successful candidate will possess a strong background in electron microscopy with an emphasis on applications to materials science. Demonstrated skills in STEM, TEM, HREM, PEELS, EDS and associated computer programs. Research experience in electron microscopy applications in materials science. Effective written and oral communication skills, as evidenced by publications and presentations in the electron microscopy field. Must be able to perform successfully in a team environment.
Desired Skills, Knowledge, Abilities:
Experience in operating an electron micoscopy facility involving large numbers of users.
Education, Training, or Licensing:
Ph.D. in materials science, physical sciences, or equivalent combination of education and experience. Minimum of 10 years experience in operating transmission electron microscopes.
Qualified people can indicate their interest by e-mailing me and attaching their cv.
Terry Mitchell
********************************** * Terence E. Mitchell * Center for Materials Science * Mail Stop K765 * Los Alamos National Laboratory * Los Alamos, NM 87545 * Tel: 505-667-0938 * Fax: 505-665-2992 * E-mail: temitchell-at-lanl.gov **********************************
I produced a sample for Ar profiles by gluing a graphite disc to a copper slot grid then dimpling and ion beam thinning it. I found that sufficient Ar was implanted into the graphite to allow suitable spectra from which profiles could be made.
} Dear Fellow Sufferers, } I am trying to analyse sputtered films by TEM/EDXA. } These contain a significant amount of Ar, but I don't have a } suitable standard (isn't that strange!). I can calculate a } theoretical k factor which will give a reasonable estimate, } but that still leaves me scratching around for a decent peak } profile. Has anyone any solutions to this problem? } Regards, } Eric } } } Dr Eric Lachowski } University of Aberdeen } Department of Chemistry } tel +44 1224 272934 } fax +44 1224 272921 } e.lachowski-at-abdn.ac.uk
Mark Blackford TEM Group Materials Division, Ansto PMB 1, Menai, N.S.W. Australia 2234 Phone (02) 9717 3027 Fax (02) 9543 7179
Gary, No camera is necessary. 1. You can use a Mac based sytem to control the SEM (4 Pi Analysis makes one; www.4pi.com) or a PC based system. these will cost US$8,000 and up. 2. some kind soul who saved it might send you a recent message outlining how to build your own Mac controller for under $1,000. 3. Run a coax cable from the framegrabber in your Mac to the video out BNC on your SEM. On our JEOL 6300, the video out is behind the kick panel under the console. This last is the simplest method, but you are limited to the resolution of the signal to the video monitor. However, I found that it is quite sufficient for many purposes.
} We are trying to find out how to connect a Mac computer to a Scannig EM } Jeol JSM 5310 microscope. Regards, Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
} Some of you know more about the Particle Atlas CD than I do. } Where is it available? Who does it? How much it costs? } The last Particle Atlas I saw (and have) was published by } Ann Arbor Science in 1973 (Second Edition). } There should be something fresher out there... } Thanks for any information I get.
} Kris } ================================================================= } Dr. Kristof KOVACS } Associate Professor } University of Veszprem, Central Laboratory } P.O.Box 158, Veszprem, HUNGARY } H-8201 } Phone: +36-(88)-421-684 } *********************************************************
Also, please -- What sorts of particles? What sizes? What microscope(s)?
:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.: Michael A. O'Keefe, Deputy Head National Center for Electron Microscopy Lawrence Berkeley National Laboratory University of California Berkeley, California 94720 tel: (510) 486-4610 fax: (510) 486-5888 email: maok-at-lbl.gov http://ncem.lbl.gov/ :.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:
Message-Id: {1.5.4.32.19961023171909.006cddf4-at-vet.purdue.edu} X-Sender: jjt-at-vet.purdue.edu X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
HI,
I am soon going to start working in a project that involves autoradiography on the electron microscopical level. There is not very much written about it (I've read Baker), so I have quite a few questions... One of my questions is regarding exposure time; I'll be working with a gamma-radiator (In-111).
If you have any idea, please contact me!
My regards,
Margareta Halin Dept. of anatomy and histology SLU Sweden
We have performed autoradiography using IN-111 on one micron sections of epoxy embedded tissues. The half-life of IN-111 was so short that within 5 days after collecting tissues from animals injected with an IN-111 compound, there was not sufficient radioactivity left to expose an emulsion. The trick is to get the material embedded, sectioned, and dipped in emulsion as soon as possible. I would suggest a rapid embedding procedure (microwave) so that you get the emulsion on the sections within 24 hours. An exposure time of 3-5 days should be sufficient. John J. Turek, Ph.D. Purdue University Dept. of Basic Medical Sciences Core Laboratory for Image Analysis and Multidimensional Applications (CRISTAL) phone: 317-494-5854 fax: 317-494-0781
The topic of artefact in microscopy goes back at least to Robert Hookes "Microscopia" in which he warns microscopists against "mistaking the shadow for the substance" Pretty apt even today.
I think the only honest way is to take the approach suggested by Edgar Mercer. All biological electron microscopy IS AN ARTEFACT. Sure, it starts with a real thing and is based on a real thing but it is not the same as the real thing. Our only excuse is that we understand the changes we have induced and that the artefact is repeatable. There are degrees of change, too. A gold coated fly in an SEM has been altered less than a sectioned chloroplast.
It is instructive to read the challenges to electron microscopy thrown out by the gadfly Harold Hillman. See Hillman and Sartory, Perception, 1977 Vol 6, pp 667-673 and anything else Harold has challenged. Get your students to read them and answer the challenges. THAT will teach them some microscopy!
The Particle Atlas Electronic Edition was put out by MicroDataware which is no longer in business. It is a very nice electronic, computer searchable copy of the 6 Volumns of the Old Particle Edition. The search ability is NICE!!!!!
The cost was about US$ 1500.00.
As I stated MicroDataware, and this is a shame, is no more. From the note I saw here, it seems that the McCrone Research Institute is looking at re-issue of it in some form.
If they do, I wish them luck. But specifically I wish them sales.
Shalom from Jerusalem, Azriel
On Wed, 23 Oct 1996, Kris Kovacs wrote:
} Dear Fellow Microscopists, } } Some of you know more about the Particle Atlas CD than I do. Where is it } available? Who does it? How much it costs? The last Particle Atlas I saw } (and have) was published by Ann Arbor Science in 1973 (Second Edition). } There should be something fresher out there... } Thanks for any information I get. } } Kris } ================================================================== } } } } ************************************************************************* } Dr. Kristof KOVACS } Associate Professor } University of Veszprem, Central Laboratory } P.O.Box 158, Veszprem, HUNGARY } H-8201 } Phone: +36-(88)-421-684 } ************************************************************************* }
} We are currently upgrading our Image analysis software and are } considering purchasing either Universal Imaging's Metamorph or } COMPIX's C*Imaging product. Dear Lloyd, We have been using the Compix product for two years now. The system is used almost all day, every day for grain size measurements on metal. It is very robust, easy to use and modify for your particular use. i have no knowledge of the Universal product. Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
This isn't my area, but I thought people had switched to various methods where the target is visualised with gold probes, because film exposure times were so long. Anyway, there is reference to several papers (almost all in animal cells) using radiolabelled probes in Hall and Hawes (1991) - Electron microscopy of Plant cells - pp. 221-222, that may be useful.
cheers,
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Clayton, Victoria 3168, Australia phone 61-3-9905 5670 email r.g.white-at-sci.monash.edu.au fax 61-3-9905 5613 or rgwhite-at-vaxc.cc.monash.edu.au
If you are in need of 10 bit/ or very high signal to noise } 62dB monochrome real time CCD cameras, digital or analog, for your microscope systems feel free to look at our very informative web site / Q &A section, and rundown on how to choose a CCD camera, and what to look for.
http://www.edt.com/dvc/dvc.html
We supply complete PCI bus imaging systems/ frame grabbers etc., from many different manufacturers in US and Canada. Software also.
Regards,
Richard Klotsche DVC Company 619-444-8300 619-444-8321-fax
To all Is there anyone who has experience of TEM immunohistochemistry on human skin? Can he send us the methods used and is observations on cryofixation, cryoprotection and so on?
Thanks you.
Prof. Paolo Castano Istitute of Anatomy Via Mangiagalli 31, 20133 Milan Italy E-mail clsmteam-at-imiucca.csi.unimi.it
What does everyone else do with their osmium tetroxide waste?
We only use a few grams a year so it is allowed to accumulate in a waste bottle of osmium 'black' until it annoys me and goes to a waste disposal company. I have worked in a lab where it was re-cycled into low grade osmium tetroxide but it was time consuming and the end product could not be used for demanding work.
Are there any commercial companies which re-process the waste or is this un-economical because of the relatively low value of osmium, at present, and the potential mix of buffer vehicle components? The reason I ask is that I used to hear rumours about chemical companies recovering osmium but I have never actually traced one.
This question is more about the environment than economics and I know the same question can be asked of other elements such as silver in photography.
Incidentally, I would like to thank everyone for their suggestions about our problem with a molybdenum artefact peak in x-ray microanalysis. We have greatly reduced the problem by having our service engineer paint the exposed edges of the objective aperture rod with carbon DAG.
Malcolm Haswell E.M Unit University of Sunderland UK
Our Pathology /Imaging group has been using the C-imaging system for 3 years and find it very useful for quantitating samples ranging from immunocytochemistry slides to autoradiograms. I have not seen an imaging product that can perform 1 Mb grey scale operations as fast as Simple with the 1280 Matrox boards (1000 mips, In was told). Software is very user friendly, and a win95 version is due out soon. I strongly recommend. If you would like more info, you can e-mail directly at davidloudy-at-mmd.com.
A friend who does SEM mostly but is doing more TEM asked me the other day why her epon blocks were sticking in the mold. She said that they were cutting them out with razor blades but it was getting costly. I also use razor blades and have cut myself numerous times when the blade stuck. I even ended up with stitches on two fingers from one of these incidents. I have heard of using liquid nitrogen to remove blocks but as I've never seen it done, I have no idea how this works. But I thought there must be a better way to remove blocks. Does anyone have any easier and safer methods for removing epon blocks from their molds?
Dozens, if not hundreds, of papers describing collodial gold lectin labeling of the extracellular membrane are out there. Among them is Frisch & Phillips 1990 Lectin binding patterns to plasmalemmal glycoconjugates of goblet cells undergoing differentiation in vitro. J. Electron Microscopy Technique 16:25-36. WGA would be an obvious starting place. Another common approach is to use cationized ferritin to non-specifically label the extracellular face of the plasmalemma. Cationized gold is also now available.
} Hi everybody, } I need some advice. } We would like to check for the proportion of vesicles which are outside } out as opposed to inside out in liver plasma membranes. } Does anyone know of, or has used, a cytochemical marking for TEM to } selectively label: } a) only the extracellular domain or } b) only the intracellular domain of a typical common plasma membrane } protein. } In particular, has anyone used colloidal gold-tagged lectins to bind } membrane glycoproteins? If so, do you have a protocol? } Any advice will be appreciated including suggested readings. } TIA } Regards, } John Gabrovsek } CCF Cleveland, Ohio
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Does anyone know of any commercial services providing sectioning and/or cryosectioning of bio materials near me in northern Calif.? Please respond by E-mail.
Thanks,
Mark A. Wall Lawrence Livermore National Lab Livermore, CA 94550 510-423-7162
Greetings Friends, Can anyone help with a proceedure for CPD a 50um free floating section? It is brain tissue that has been fixed, cryosectioned and returned to buffer. All wizardry is welcome at this point !! Has anyone used Peldri II ? Would this be a possibility? Thanks Linda Fox lfox1-at-wpo.it.luc.edu
In the spring a European journal asked for labs that wanted to publish their abilities space in a North American listing of microscopic labs. We sent in a listing and recieved a phone call from England about viewing the listing. The number that we got off the answering machine keeps giving us an out of order number. If any one has the number then please share it with us. Thank you, Clarissa Vierrether ORES 1200 West Third Salem, MO 65560 cvierret-at-misn.com
I am a graduate student at NC State University in an SEM class. I am currently performing a research project on the development and history of the scanning transmission electron microscope (STEM). I was wondering if someone could give me an idea of what break- throughs have been made over the past few decades and possibly what references I could look for that might concern these breakthroughs.
Mr-Received: by mta RANDD; Relayed; Thu, 24 Oct 1996 08:21:51 -0500 Mr-Received: by mta MCM$RAND; Relayed; Thu, 24 Oct 1996 08:21:52 -0500 Mr-Received: by mta RANDD; Relayed; Thu, 24 Oct 1996 08:22:03 -0500 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
Brad,
We have a copy of the Particle Atlas on CD-ROM and use it fairly regularly. We also do a lot of light microscopy and electron microscopy similar to what was done in the atlas. I do wish there was away to add images and data to the atlas that are more typical of the types of particles we encounter. This feature would make it an extremely valuable resource but would likely change the atlas into a subscription. Either way we felt it was worth the price we paid which I think was $950 (we got a show special from the Intermicro meeting in '95). It would also be nice if it were available in a Mac version.
Joe Neilly Abbott Laboratories Abbott Park, IL 60064
Has anyone done any microscopy work on alpha Pb? This is lead that has low alpha radiation and is useful in the electronics industry. We have some samples of "regular" lead and low alpha lead. WE would like to know if there is a way to distingush between the two microscopically. The alpha emissions test are very costly and seem to have a delayed turn around time. Please contact me directly at cvierret-at-misn.com. I will post the answers if there are any. Thank you for your time, Clarissa
I am looking for some STM and AFM pictures to show to our students within the frame of an electron miroscopy course. I intend to tell them that there are also other microscopies than the classical light microscopy and the TEM/SEM. I would also point out how pictures can be misleading if you don't know anything about the technique used to get them and about their scale.
In one example, I am trying to get as much as possible images of dislocations of any kind (vein on saguarro cactus, painting on wall, mosquito's eye, TEM=8A). If by any chance do you have a scanning probe image of a (edge) dislocation observed at the atomic/molecular scale on metals, ceramics, semi-conductors, or on a biological cristal, would you accept to send me a copy (a file by e-mail would be fine and probably the easiest, a print on paper would also be nice).
Of course, any other pictures are also welcome.
Thanks in advance for your help and time. Best regards
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
Message-Id: {199610241827.NAA09315-at-Sparc5.Microscopy.Com} To: Microscopy messageslistserver {Microscopy-at-Sparc5.Microscopy.Com}
We don't usually have this problem, since we choose our molds with proper release in mind. Polyethylene, as in BEEM products, give the easiest release. Silicon rubber is also good and can be made better by spraying with a release agent. Polypropylene can also be used and the epon will release with mechanical pressure as with a hammer or a pair of pliers or with a hacksaw. If you are using polstyrene, then you have troubles. Here is where dipping in liquid nitrogen might help, although you have to hope that your epon does not shatter right through the important part of the specimen. Selection of mold material is the secret.
A friend who does SEM mostly but is doing more TEM asked me the } other day why her epon blocks were sticking in the mold. She said that } they were cutting them out with razor blades but it was getting costly. I } also use razor blades and have cut myself numerous times when the blade } stuck. I even ended up with stitches on two fingers from one of these } incidents. I have heard of using liquid nitrogen to remove blocks but as } I've never seen it done, I have no idea how this works. But I thought } there must be a better way to remove blocks. Does anyone have any easier } and safer methods for removing epon blocks from their molds? } } Thanks in advance!! } } Lesley Bechtold } } } ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
} Date: 24 Oct 1996 08:07:49 -0800 } From: Mark Wall {Mark.Wall-at-quickmail.llnl.gov} } To: List server {microscopy-at-Sparc5.Microscopy.Com} } Subject: Mark Wall } } Does anyone know of any commercial services providing sectioning and/or } cryosectioning of bio materials near me in northern Calif.? Please respond by } E-mail. } } Thanks, } } Mark A. Wall } Lawrence Livermore National Lab } Livermore, CA 94550 } 510-423-7162 } Hi, We do lots of sectioning and ultracryosectioning for lots of people. Although we're not in No. CA, there is FedEX. We receive samples by FedEx from all over for electron microscopy diagnostic virology (can provide details on this too if anyone is interested). There's no reason why you couldn't send cells/tissues in glutaraldehyde or paraformaldehyde for processing. Let me know if we can help.
Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Date: 24 Oct 1996 08:07:49 -0800 } From: Mark Wall {Mark.Wall-at-quickmail.llnl.gov} } To: List server {microscopy-at-Sparc5.Microscopy.Com} } Subject: Mark Wall } } Does anyone know of any commercial services providing sectioning and/or } cryosectioning of bio materials near me in northern Calif.? Please respond by } E-mail. } } Thanks, } } Mark A. Wall } Lawrence Livermore National Lab } Livermore, CA 94550 } 510-423-7162 } Hi, We do lots of sectioning and ultracryosectioning for lots of people. Although we're not in No. CA, there is FedEX. We receive samples by FedEx from all over for electron microscopy diagnostic virology (can provide details on this too if anyone is interested). There's no reason why you couldn't send cells/tissues in glutaraldehyde or paraformaldehyde for processing. Let me know if we can help.
Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Forwarded message: } From KCH7-at-cornell.edu Wed Oct 23 20:21:30 1996 } Message-Id: {2.2.32.19961024002431.00694f4c-at-postoffice4.mail.cornell.edu} } X-Sender: kch7-at-postoffice4.mail.cornell.edu } X-Mailer: Windows Eudora Pro Version 2.2 (32) } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Date: Wed, 23 Oct 1996 20:24:31 -0400 } To: John Hunt {hunt-at-msc.cornell.edu} } X-UIDL: 846158903.012 } From: "Kenneth C. Hover" {KCH7-at-cornell.edu} } Subject: Re: Quantitative microscopy of packed particles (fwd) } } John: } } One of my colleagues at NIST has worked in this area. His name is Ken } Snyder, working in the Building Materials Lab under Dr. Geoff Frohnsdorff. } I am not in my office but can get address and email. } } ken } } } } } } At 09:41 AM 10/16/96 -0400, you wrote: } } Forwarded message: } } } From Microscopy-request-at-Sparc5.Microscopy.Com Tue Oct 15 15:42:36 1996 } } Message-Id: {s263847e.073-at-intergate.dot.gov} } } X-Mailer: Novell GroupWise 4.1 } } Date: Tue, 15 Oct 1996 12:32:16 -0400 } } From: "N. Shashidhar" {NSHASHIDHAR-at-intergate.dot.gov} } } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: Quantitative microscopy of packed particles } } } } I need to quantify pavements, which can be looked at as a system of packed } particles. I need to } } obtain the number of contacts each particle has on average with another } particle. This can then be } } related to mechanical properties such as shear resistance, etc. } } } } Are there microscopic techniques to get this information from a compact and } stereology methods to } } extract such information. Any response will be appreciated. } } } } Thank You. } } } } } Kenneth C. Hover, P.E., Ph.D. } Associate Dean for Undergraduate Programs } } 221 Carpenter Hall } Cornell University } Ithaca, New York 14853 } } Phone 607-255-0393/8340 } Fax 607-255-9606 } Email kch7-at-cornell.edu }
I have a Postdoc. going over to Holland for a six month placement. She wants to know where to find a reliable method to embed tissue that contains ceramic or metallic implants - she hopes to preserve a number of bone proteins in the process (Vitronectin, Fibronectin & Osteopontin) for immuno. work in 1997. The questions are:
1. Do we have to use a cryo/freeze-sub. route to preserve this tissue, or might we get away with a "light" glut. fixation?
2. Would LR Gold be an appropriate resin, bearing in mind me may want to do LM and TEM?
Finally, does anybody know a source for commercially pure titanium rod (diameter 1.5 to 2.0 mm)?
Thank you for your help. Paul
Dr Paul V. Hatton Lecturer in Biomaterials School of Clinical Dentistry University of Sheffield Claremont Crescent SHEFFIELD S10 2TA
Tel. (0114) 271 7938 Fax. (0114) 2665326 or 2797050
Alternate-Recipient: allowed Auto-Forwarded: prohibited Content-Return: allowed Disclose-Recipients: prohibited Conversion: allowed Importance: normal Priority: normal Sensitivity: Company-Confidential Microscopy-at-Sparc5.Microscopy.Com
Hi,
Not too long ago, I put up a homepage for a university open house (, thus not designed for academic purpose,) which may have part of what you need. The pages show how various scales can make a difference to the microstructures observed. You can find these at:
http://mse.mcmster.ca/research/micro
Suggestions are welcome as well.
============================================================================ J. K. Chen, PhD | Postdoctoral Fellow 357 Engineering Bldg. | E-mail: jkchen-at-mcmaster.ca McMaster University | Phone: +1-905-5259140 ext.27042 (O) Dept. of Materials Sci. & Eng.| Fax: +1-905-5289295 Hamilton, ON L8S 4L7, CANADA | http://mse.mcmaster.ca/faculty/cv/jkchen ============================================================================
At 04:16 PM 10/24/96 +0100, you wrote to Microscopy listserver: } ... snip ... } I would also point out how pictures can be misleading if you don't } know anything about the technique used to get them and about their scale. } ... snip ... } } Of course, any other pictures are also welcome. } } Thanks in advance for your help and time. Best regards } } Philippe Buffat
} Greetings Friends, Can anyone help with a proceedure for CPD a } 50um free floating section? It is brain tissue that has been fixed, } cryosectioned and returned to buffer. All wizardry is welcome at } this point !! Has anyone used Peldri II ? Would this be a } possibility? Thanks } Linda Fox lfox1-at-wpo.it.luc.edu
Linda; CPD: cut the end off of a BEEM capsule, cut out the center of the capsule's cap, cut off the cap of another BEEM capsule, cut out the center, put plankton netting over the end of the capsule with the lid, put in sections, close lid; dehydrate, standing open end up in a Wheaton 4 mL vial, add EtOH to capsule, withdraw from vial outside of capsule; put on cut-off lid with netting, put in CPD & dry:
Peldri: fluorocarbon, not made anymore. or: dehydrate as usual, and dry from HMDS (Hexamethyldisilizane). Phil
&&&&&&&&&&& Illigitmi non carborundum &&&&&&&&&&&
Philip Oshel *all mail:* Microscopy University of Illinois Station A Rm 74 Bevier Hall PO Box 5037 905 S. Goodwin Ave. Champaign, IL 61825-5037 USA Urbana, IL 61801 (217) 244-3145 oshel-at-ux1.cso.uiuc.edu (217) 355-1143 home
At 1:16 PM 10/24/96, cvierret-at-misn.com wrote: } Has anyone done any microscopy work on alpha Pb? This is lead that has low } alpha radiation and is useful in the electronics industry. We have some } samples } of "regular" lead and low alpha lead. WE would like to know if there is a way } to distingush between the two microscopically. The alpha emissions test are } very costly and seem to have a delayed turn around time.
WHY COSTLY ? Alpha is easy to detect and assay, yes ?
510-429-1060 Fax 429-1065 LMDC, (Laser & Motion Development Co.) 3101 Whipple Road Union City, CA 94587-1216
TO BE ON OUR LIST AND BE THE FIRST TO KNOW ABOUT BARGAIN PRICED TECHNOLOGY EQUIPMENT, Just send an e-mail containing "Join Equipment List" in the header line.
Our web page: http://www.lasermotion.com Our e-mail: office-at-lasermotion.com
} } What does everyone else do with their osmium tetroxide waste?
} For a few years I coordinated a scheme in Australia where anyone could send me their Osmium waste. We precipitated it by adding formaldehyde, I stored it until we had about 100 litres then we sent it to Johnson Matthey. They filtered off the osmium precipitate and I think then sent the cake to Matthey in the UK for refining. The recovery rate was much less than we hoped (probably due to the volatile nature of the oxides) and out of the value of the osmium that was recovered, most of the money went to pay the refining cost. All we were left with was a warm fuzzy feeling that we had helped recycled a scarce resource.
But do try it if you like. Contact MAtthey in the UK (They may be Matthey Garrett, I cant track all these company name changes) as they certainly know about refining osmium.
Message-Id: {1.5.4.32.19961025012758.006a6384-at-mailhost.ultra.net.au} X-Sender: pns-at-mailhost.ultra.net.au X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
At 16:16 24/10/96 +0100, you wrote: } Dear Dieter, } } I am looking for some STM and AFM pictures to show to our students within } the frame of an electron miroscopy course. I intend to tell them that there } are also other microscopies than the classical light microscopy and the } TEM/SEM. I would also point out how pictures can be misleading if you don't } know anything about the technique used to get them and about their scale. } } In one example, I am trying to get as much as possible images of } dislocations of any kind (vein on saguarro cactus, painting on wall, } mosquito's eye, TEM=8A). If by any chance do you have a scanning probe= image } of a (edge) dislocation observed at the atomic/molecular scale on metals, } ceramics, semi-conductors, or on a biological cristal, would you accept to } send me a copy (a file by e-mail would be fine and probably the easiest, a } print on paper would also be nice). } } Of course, any other pictures are also welcome. } } Thanks in advance for your help and time. Best regards } } Philippe Buffat } } __________________________________________________________________ } Philippe Buffat } Ecole Polytechnique Federale de Lausanne (EPFL) } Centre Interdepartemental de Microscopie Electronique } Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland } Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) } E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ } ______________________________ Eudora F2.1 ___________________________ Hello Philippe and whoever: Within our on-line catalogue is an excellent collection of microscopy related links. You will find numerous images to chose from in the various Atomic/Tunnelling microscopy sites and endless more microscopy images under the header "Image Resources". I expect that you know that you can copy images to disk by clicking on the right mouse button. Images on the internet are covered by copyright but you could ask to use these and many posted images in fact specifically permit their use for teaching purposes.= =20 Cheers Jim Darley =20 Probing & Structure } (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } A great microscopy site http://www.ultra.net.au/~pns/
Message-Id: {199610250402.AA09220-at-lucy.swin.edu.au} Comments: Authenticated sender is {hbrinkies-at-gpo.swin.edu.au}
I know this server is mainly concerned with electron microscopy but I need some information on optical equipment.
As most of us know university funds are getting constantly smaller. I am therefore trying to refurbish am older Leitz micro-hardness tester that had been stored in a cupboard for several years. The objectives are badly damaged. Where can I find replacements ? The objectives are marked A 0.18 C HM 25oo 10x and A 0.70 C HM 6.3 40x Ernst Leitz-Wetzlar respectively.
Any help is appreciated.
Hans Brinkies SWINBURNE, University of Technology School of Mechanical and Manufacturing Engineering Industrial Microscopy HAWTHORN, 3122, Australia
There was a short 'Technical Tip' in the procedings of the RMS some time ago re. osmium disposal which I took to heart! I bring in old cooking oil and tip it into that so that it becomes bound and inactivated. When there is enough, I mix it with a hot water / detergent mixture (Quadralene Instrument Cleaner) and then wash down the sink with 'copious amounts of running water'. No matter what you do with it, it has to end up in the environment from whence it came?
With best wishes - Keith Ryan
++++++++++++++++++++++++++++++++++++++++++++++++++ Keith Ryan Plymouth Marine Laboratory, Citadel Hill, Plymouth, Devon PL1 2PB, England
To avoid cut fingers when slicing open embedding capsules I use a TAAB capsule splitter. This is a metal block with a hole drilled at either end to take a 6 or 8mm capsule and two slits on either side to enable a single-edge razor blade to be inserted and drawn down the side of the capsule without the risk of it slipping and slicing into your fingers. The nearest distributor to you would be in Canada at Marivac Ltd, 5821 Russel Street, Halifax, Nova Scotia, B3K 1X5. It costs 26.72 UK pounds.
Dan Hill Biochemistry Dept Cambridge University United Kingdom
On Thu, 24 Oct 1996, Lesley S. Smith wrote:
} A friend who does SEM mostly but is doing more TEM asked me the } other day why her epon blocks were sticking in the mold. She said that } they were cutting them out with razor blades but it was getting costly. I } also use razor blades and have cut myself numerous times when the blade } stuck. I even ended up with stitches on two fingers from one of these } incidents. I have heard of using liquid nitrogen to remove blocks but as } I've never seen it done, I have no idea how this works. But I thought } there must be a better way to remove blocks. Does anyone have any easier } and safer methods for removing epon blocks from their molds? } } Thanks in advance!! } } Lesley Bechtold } }
I am a user of a JEOL6400 SEM with LaB6 gun. I have noticed that there is a new cathode on the market called DENKA LKS7. Has anyone got any experience in using this type of cathode.
Message-Id: {1.5.4.32.19961025131001.0069f74c-at-biotech.ufl.edu} X-Sender: sdw-at-biotech.ufl.edu X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Have you tried Hexamethyldisilizane (HMDS for short). We have found it more convient than Peldri and it gives similar to better results. There are discussions about both which came across the list and I have archived them at the web site listed at the end of this message. Click on the "Tips & Tricks " button and look in the SEM section. Let me know if you cannot access it and I will get it to you some other way
At 10:10 AM 10/24/96 -0500, you wrote: } Greetings Friends, Can anyone help with a proceedure for CPD a } 50um free floating section? It is brain tissue that has been fixed, } cryosectioned and returned to buffer. All wizardry is welcome at } this point !! Has anyone used Peldri II ? Would this be a } possibility? Thanks } Linda Fox lfox1-at-wpo.it.luc.edu } } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Your comment on no matter what you do with it, it has to end up from whence it came sounds like what the big manufacturing companies used to say when they were found dumping large volumes of untreated waste into rivers and streams. These things are no longer in their natural form and so they can do more harm. I like the idea of recycling even if you only break even. At least you know it won't be showing up in your drinking water or food in the near future...That's my two cents.
Karen
On Fri, 25 Oct 1996, Keith Ryan wrote:
} Dear Malcolm } } There was a short 'Technical Tip' in the procedings of the } RMS some time ago re. osmium disposal which I took to } heart! I bring in old cooking oil and tip it into that so that it } becomes bound and inactivated. When there is enough, I } mix it with a hot water / detergent mixture (Quadralene } Instrument Cleaner) and then wash down the sink with } 'copious amounts of running water'. No matter what you do } with it, it has to end up in the environment from whence it } came? } } With best wishes - Keith Ryan } } ++++++++++++++++++++++++++++++++++++++++++++++++++ } Keith Ryan } Plymouth Marine Laboratory, Citadel Hill, } Plymouth, Devon PL1 2PB, England } } Tel: ++44 1752 633294 (international) } 01752 633294 (national) } Fax: ++44 1752 633102 (international) } 01752 633102 (national) } e-mail: k.ryan-at-pml.ac.uk } PML web site: http://www.npm.ac.uk/pml } ++++++++++++++++++++++++++++++++++++++++++++++++++ } }
I would like very much to thank all those who responded to my inquery/plea for help/ideas on placement and in house support for university wide em facilities. The ideas were gobbbled up by our newly appointed Users Committe. We are putting together our recommendations at the end of next week to the central adminstration and then cross our fingers and wait. Thanks Again, Hank Adams EML New Mexico State University Las Cruces, NM "The Green/Red Chile Capitol"
Could you please place the following ad on the Microscopy Server? Please let me know that you got this message.
Thank you. ***************************************************************** SENIOR FACULTY POSITION IN MICROSCOPY/MICROANALYSIS/ MATERIALS SCIENCE AND ENGINEERING
The Department of Materials Science and Engineering at Lehigh University has an opening for a senior (associate/full) professor with expertise in electron microscopy and/or microanalysis (SEM, TEM, AEM, etc.). The successful candidate must have a proven record of research in the application of microscopy and/or microanalysis to the solution of materials problems and be able to conduct independent and cooperative research in MS&E. Ability to teach undergraduate and graduate courses in microscopy and materials is essential. Experience in running a microscopy facility will be an advantage. Curriculum vitae and the names of three references should be sent by December 15, 1996 to:
Professor David B. Williams Chairman, Search Committee Dept. of Materials Sci. & Eng. Lehigh University 5 East Packer Avenue Bethlehem, PA 18015-3195
Lehigh University is an committed to recruiting, retaining, and tenuring women and minorities. ***************************************************************************
Sharon L. Coe Department of Materials Science & Engineering 5 East Packer Avenue Lehigh University Bethlehem, PA 18015 610/758-5133 e-mail: slc6-at-lehigh.edu
Dear Group, I have been asked to give a couple of lectures in February as part of an interdisciplinary lecture course on 'The new microscopies'. I will be discussing HREM, and since the lectures are aimed at both physical and biological scientists I would very much like to include some life science examples. I know that very good work has been done using cryo-HREM. If anybody has any images that I would be able to use (suitably referenced of course) or could suggest people to contact, I would be most grateful.
Yours, Amanda Petford-Long
Dept. of Materials, Univ. of Oxford Parks Road, Oxford OX1 3PH, UK Tel +44 1865 273656 Fax +44 1865 283333
The simplest way to avoid cuts while opening BEEM capsules is to set the capsule upright on the bench, resting on its flat, uncapped end. Then, perhaps with the middle finger of each hand supporting the capsule on the side away from you, slide a razor blade down the near side. But, hold the razor blade tangentially to the capsule. Removing a broad strip of plastic make the thing easier to open and dump the block out, and allows better control over the blade motion. } } On Thu, 24 Oct 1996, Lesley S. Smith wrote: } } } A friend who does SEM mostly but is doing more TEM asked me the } } other day why her epon blocks were sticking in the mold. She said that } } they were cutting them out with razor blades but it was getting costly. I } } also use razor blades and have cut myself numerous times when the blade } } stuck. I even ended up with stitches on two fingers from one of these } } incidents. I have heard of using liquid nitrogen to remove blocks but as } } I've never seen it done, I have no idea how this works. But I thought } } there must be a better way to remove blocks. Does anyone have any easier } } and safer methods for removing epon blocks from their molds? } } } } Thanks in advance!! } } } } Lesley Bechtold } } } }
Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, Wa 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
In response to Scott's suggestion that an Ar standard could be made, it seems to me that if you know your detector well enough and can determine good k-factors for K and Cl, then you would be better off estimating the ArX k-factor than trusting that you've properly manufactured and estimated the concentration of the *standard*. DTSA is a program that would be helpful in this situation. It takes some effort to fully characterize the efficiency of your detector and then choose which set of empirical parameters work in your analytical environment (e.g., ionization cross sections, etc.), but it has been worthwhile for our lab. Ion implanting of Ar in C (or one of the sheet silicate minerals like pyrophyllite) could give you the shape reference. Good luck.
Ciao for now, Ken
} I was following this topic because I once looked at some ion sputtered } hydroxyapatite films that had Ar incorporated in them. I ignored it because I } figured it would diffuse out eventually. } } Mark Blackford's answer gave me an idea for an approach to the solution to this } problem. It is based on low energy ion implantation. The problem with Mark's } answer is that you do not know the concentration of the Ar in the graphite from } the ion milling process and therefore cannot calculate the k-factor. You would } also be calculating the k(C,Ar) factor and would be tough to correlate it with } other k(C,X) factors since carbon is tough to quantify in a lot of materials. } Even though the graphite is tough to ion mill with straight Ar you will still } lose C while you mill. You need to use a material that resists ion milling } from Ar and was not prepared by ion milling. I suggest using W. The W could } easily be electrolytically prepared with 5% NaOH solution (5V ac) and the TEM } foils implanted with Ar at about 1 keV. (It would be better to prepare the W } with a known geometry such as would result by Tripod Polishing.) The } concentration profiles could be calculated using some of the Monte Carlo TRIMML } codes or other codes that are available. Unfortunately, they are not great at } low energies. Experimental alternative are to do depth profiling of implanted } field emitters in an atom probe(Been there, done that.), SIMS profiles, or RBS } profiles. If you know the thickness of the W sample and the concentration } profile of the implanted Ar, you could calculate the concetration of the Ar in } the volume probed in the AEM. You could then calculate the k-factors from the } measured intensities. A benefit of using W is that you can work with } relatively thick samples without worrying too much about the thin film } criteria. You are going to have large errors with this approach. } } Other materials that might work would be thin films of Au or Au/Pd that were } ion implanted. } } This is not an easy problem. These are some thoughts off the top of my head, } so you should take them as such. } } - -Scott Walck }
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets The Johns Hopkins University Baltimore, Maryland 21218
I would like to be in touch with someoen in the S.F. Bay area that is doing freeze-fracture electron microscopy and could do ultrarapid freezing of uncryoprotected sample. I have a collaborator there and it would be easier if he could work with some one locally.
Reply directly and not to the server.
Thank you/. ******************************************************* Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
The best technique for this would likely be xray tomography. One can obtain 3d images of samples with 1 to 10 um resolution. Element-specific maps can also be made, so one could obtain both morphological & elemental info in 3d. Performing such experiments is best done at a dedicated facility (beamlines; such as Brookhaven or NIST). Depending upon the analysis needs though, bench-scale measurements using more standard xray generators can produce very useful images.
Once the images are in hand, one simply uses standard image processing techniques to extract the desired parameters (contact points, contact areas, angles, etc.).
Good luck.
Regards, Bill.
} Subject: Quantitative microscopy of packed particles } } I need to quantify pavements, which can be looked at as a system of } packed particles. I need to obtain the number of contacts each } particle has on average with another particle. This can then be related } to mechanical properties such as shear resistance, etc. } } Are there microscopic techniques to get this information from a } compact and stereology methods to extract such information. Any } response will be appreciated. } } Thank You. } } } Kenneth C. Hover, P.E., Ph.D. } Associate Dean for Undergraduate Programs } } 221 Carpenter Hall } Cornell University } Ithaca, New York 14853 } } Phone 607-255-0393/8340 } Fax 607-255-9606 } Email kch7-at-cornell.edu William A. Lamberti Office LA-196 Exxon Research and Engineering Company Route 22 East Annandale, NJ 08801 (908)730-2144 office (908)730-2262/2104 labs (908)730-3042/3051 fax Email: walambe-at-erenj.com
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At 09:58 AM 10/25/96 -0400, you wrote: } In response to Scott's suggestion that an Ar standard could be made, it } seems to me that if you know your detector well enough and can determine } good k-factors for K and Cl, then you would be better off estimating the } ArX k-factor ...
This suggestion reminds me of an ancient method of synthesizing peaks given only a few measured spectra. The idea was, for a given keV and detector low-energy efficiency, acquire a spectrum of pure carbon (e.g., from polished vitreous carbon or diamond) and assume this spectrum defines the envelope (shape) for all spectral peaks for the pure elements. Kenneth's suggestion of measuring K and Cl, together with the C "efficiency" spectrum would provide excellent criteria for knowing exactly what the Ar intensity should be.
cheers, shaf
{\/} /\ {\/} /\ {\/} /\ {\/} cognito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
I dont remember epon sticking to polythene or polystyrene if it was fully cured. But epoxies will stick to silicone rubber molds when they get older. Seems that silicone rubber molds are not fully cured so thye have reactive groups free and you start to get the epoxies cross linking to the rubber. Once that happens you have to throw out the silicone rubber mold and buy (or cast) a new one.
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} Date: Thu, 24 Oct 1996 10:10:41 -0500 } From: Linda Fox {lfox1-at-wpo.it.luc.edu} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: CPD free floating section } } Greetings Friends, Can anyone help with a proceedure for CPD a } 50um free floating section? It is brain tissue that has been fixed, } cryosectioned and returned to buffer. All wizardry is welcome at } this point !! Has anyone used Peldri II ? Would this be a } possibility? Thanks } Linda Fox lfox1-at-wpo.it.luc.edu } ************************************** Linda & You: Sorry Philip O. but Peldri II is available. (Ted Pella Inc & P&S) Those compounds do nasty things to the Ozone layer but, I am afraid, my motorcar alone could produce enough nitrous oxides to exceed the damage done by all the Peldri in the world. The technical issue is the fragillity and the preservation of the section for this Peldri II would be may first choice. Jim Darley
Probing & Structure } (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } A great microscopy site http://www.ultra.net.au/~pns/
Do you know of an established method for casting molds for epoxies. I need to embed odd sized samples and have not been able to locate the appropriate sized molds so your idea of casting a mold sounds interesting!
Thanks, Louie Kerr
At 12:29 PM 10/26/96, Dr Mel Dickson wrote: } I dont remember epon sticking to polythene or polystyrene if it was fully } cured. But epoxies will stick to silicone rubber molds when they get older. } Seems that silicone rubber molds are not fully cured so thye have reactive } groups free and you start to get the epoxies cross linking to the rubber. } Once that happens you have to throw out the silicone rubber mold and buy (or } cast) a new one. } } mel dickson
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
} Mel, } } Do you know of an established method for casting molds for epoxies. I need } to embed odd sized samples and have not been able to locate the appropriate } sized molds so your idea of casting a mold sounds interesting! } } Thanks, } Louie Kerr
Hi Louie,
The easy way to cast a mold is to use silicone encapsulating rubber. We used RTV E mould making rubber we bought from a plastic supplies shop here. You make replicas exactly the shape of the final cast you want, then seal them (say with superglue) to the bottom of a shallow dish (a glass Petri dish is ideal. The replicas should have a smooth surface. You can punch numbers or letters on them if you want. Mix up the RTV resin as per instructions, degas in a vacuum desiccator and pout it into the dish and leave it to cure again as per instructions.
The silicone eventually becomes hard, brittle and adheres to the epoxies so be ready to make more molds as this happens.
Incidently, if you want to see through the roughened surface of an embedment, just smear a thin film of immersion oil over it.
The following is the text of a advertisement which will appear in next weekUs issue of the New Scientist magazine. I would be grateful if you could bring this to the attention of any of your students or colleagues who might be interested in applying for this post.
Thank you for you help in this matter, Mark Aindow
THE UNIVERSITY OF BIRMINGHAM School of Metallurgy & Materials
Research Fellow
The Nucleation Stage of Carbon Deposition on Oxidised Stainless-Steel Surfaces
Applications are invited for the above industrially funded post which will be available for two years. The work will involve producing carbon deposits on oxidised stainless-steel surfaces from carbon monoxide/dioxide mixtures and characterising them using high resolution techniques including TEM and AFM. Applicants should have a PhD and experience of relevant experimental techniques. Starting salary will be in the range 14,317 - 15,986 pounds sterling per annum.
Preliminary enquiries should be directed to: Dr. M. Aindow Telephone: 0121 414 5188, Email: M.Aindow-at-bham.ac.uk.
Application forms (returnable by 25 November 1996.) and further particulars are available from the Director of Staffing Services, The University of Birmingham, Edgbaston, Birmingham B15 2TT, Telephone 0121 414 6483 (24 hours), (Email: Staffing-at-bham.ac.uk).
Mark Aindow, School of Metallurgy and Materials, Telephone; (0121) 414 5188 The University of Birmingham, FAX; (0121) 414 5232 Elms Road, Edgbaston, Birmingham, Email; M.AINDOW-at-BHAM.AC.UK GB B15 2TT, United Kingdom.
Dear Users Group, We have a researcher interested in using Safranin O to dye bone/ cartilage sections embedded in Epon and cut at 4 microns. Has anyone done this successfully, and if so how? Any help would be appreciated.
I had a question concerning the imaging of coated and non-coated polymer fibers in scanning electron microscopy. If you could give me any useful websites and/or journal articles that are related to this topic it would be greatly appreciated. Susan Lloyd salloyd-at-eos.ncsu.edu
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Hello,
Often I am required to determine how clean a sample of strip steel is by comparing the amount of oxide remaining on the surface to the area scanned on SEM. The method we currently use is extremely time-consuming and uses up many polaroid films. Does anyone know of a simple way of finding the ratio of oxide left on a surface to bare metal? Is there any software available that may be of help?
Please reply to the group. I would also find these references interesting. (I know from some work at the light level on fixed tissue that there is often a ton of nonspecific staining in the stratum corneum. Fortunately I was interested in the dermis. But how do people get around this?)
Karen
} To all } Is there anyone who has experience of TEM immunohistochemistry on human skin? } Can he send us the methods used and is observations on cryofixation, } cryoprotection and so on? } } Thanks you. } } } Prof. Paolo Castano } Istitute of Anatomy } Via Mangiagalli 31, 20133 Milan } Italy } E-mail clsmteam-at-imiucca.csi.unimi.it }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Life Sciences Sector Lab Reply: kszaruba-at-mmm.com 3M Company 3M Center 270-1S-01 Phone: 612-737-2971 St. Paul, MN 55144-1000 Fax: 736-1519
These opinions are my own and may not represent those of 3M.
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Lesley & Glen -
Lesley didn't say specifically that she was using BEEM capsules. However, if that's the case, I recommend the BEEM Capsule Press (E.F. Fullam, Inc. cat.no. 54090; Structure Probe, Inc. cat.no. 2400-AB; Look in other suppliers' catalogs under "capsule, embedding", or "BEEM".) We produce a lot of blocks and this gizmo saves time as well as wear & tear on blocks and fingers.
Joiner
At 09:56 AM 10/25/96 -0200, you wrote: } The simplest way to avoid cuts while opening BEEM capsules is to set the capsule } upright on the bench, resting on its flat, uncapped end. Then, perhaps with the } middle finger of each hand supporting the capsule on the side away from you, } slide a razor blade down the near side. But, hold the razor blade tangentially } to the capsule. Removing a broad strip of plastic make the thing easier to open } and dump the block out, and allows better control over the blade motion. } } } } On Thu, 24 Oct 1996, Lesley S. Smith wrote: } } } } } A friend who does SEM mostly but is doing more TEM asked me the } } } other day why her epon blocks were sticking in the mold. She said that } } } they were cutting them out with razor blades but it was getting costly. I } } } also use razor blades and have cut myself numerous times when the blade } } } stuck. I even ended up with stitches on two fingers from one of these } } } incidents. I have heard of using liquid nitrogen to remove blocks but as } } } I've never seen it done, I have no idea how this works. But I thought } } } there must be a better way to remove blocks. Does anyone have any easier } } } and safer methods for removing epon blocks from their molds? } } } } } } Thanks in advance!! } } } } } } Lesley Bechtold } } } } } } } } Glen MacDonald } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, Wa 98195-7923 } (206) 616-4156 } glenmac-at-u.washington.edu } } }
On 10/28/96 11:44:34 you wrote: } } Hello, } } Often I am required to determine how clean a sample of strip steel is by } comparing the amount of oxide remaining on the surface to the area scanned } on SEM. The method we currently use is extremely time-consuming and uses up } many polaroid films. } Does anyone know of a simple way of finding the ratio of oxide left on a } surface to bare metal? Is there any software available that may be of help? } } Thanks in advance, } } Tania Jones }
Dear Tania: There is an ASTM standard protocol for QA/QC analysis of the surface oxide layer on electropolished stainless steel using SEM, Auger and XPS. It reports density of defects, average thickness of oxide layer (in Angstroms), and Cr/Fe ratio in oxide. Some procedures in this protocol could be applied to your samples. We provide SEM/EDS,Auger,SIMS,TEM,GCMS,FTIR,HPLC,ICP-MS analytical services. Igor Ivanov RJ LeeGroup 530 McCormick Str. San Leandro, CA 94577 510-567-0480 ph. 510-567-0488 FAX
I'm sorry to join the discussion late, but I haven't checked my email for about a week.
I didn't see any replies recommending the micro-chanell plate as a solution to the low kEv situation. My last impression of these systems was that they were state of the art. They work very much like a photo-multiplier, and provide the high gain and very good signal to noise ratio.
If one were seeking a BSE system for low accellerating voltage situations, it would be worth contacting GW electronics in Norcross, GA. I have no interest in GW, other than seeing that the best technologies be evaluated.
One comment about the micro-chanell: the last time I looked, they didn't offer a topographic contrast mode (neither does the robinson). Why doesn't someone build a micro-chanell plate with 2 or 4 separate detector elements and high voltage supplies so the signals can be fed into a traditional amplifier which offers atomic number and topographic contrast? Sort of a multi-detector version of the micro chanell. Is there a practical reason this can't be done, or is it a commercial reason?
---------------------------------- Dr. Wayne Lanier 250 Ashbury San Francisco, CA 94117 [415] 346-4940 lanier-at-slip.net ---------------------------------
} Hello, } } Often I am required to determine how clean a sample of strip steel is by } comparing the amount of oxide remaining on the surface to the area scanned } on SEM. The method we currently use is extremely time-consuming and uses up } many polaroid films. } Does anyone know of a simple way of finding the ratio of oxide left on a } surface to bare metal? Is there any software available that may be of help? } } Thanks in advance, } } Tania Jones } } tania-at-dynamotive.com
Don't quite understand the problem - I'd have thought that any cleaned steel surface would immediately oxidise on exposure to air, if only to a monolayer.
I guess you're talking about a fairly thick corrosion-type of oxide layer? For a reasonably flat surface, how about either an EDX map on the O-K peak, or z-contrast image via BSE (you'd probably need to adjust HT to optimise contrast depending on thickness of oxide layer)
} I have a question about standardless semi-quant work that I am doing, } involving take off angle. I have a TN5500 EDX instrument. When I analyze
snips
} My problem is, how do I perform standardless semi-quant work on the } cubes within my powder, to examine homogeneity within the sample from one } cube to the next? } } Thank You in advance. } Mark Darus } General Electric
First, I'd try running the quant programme on the same spectrum a number of times but with different values for the relevant angles. This will give you a feel for how sensitive your quant algorithm and particular combination of elements is to these paramenters.
You might find that for sample tilts over a certain value (sensitivity to specimen tilt reduces as the tilt increase - at high tilt, a small change in tilt will have less effect on X-ray path lengths, so the absorption correction will be changing less), the error is acceptably small - in which case you can choose cubes appropriately. That is, select/set up high (but not too high) tilt surfaces to analyse. I don't suppose the tilt angles of the cubes will be determined by composition. (At this point you can also check how good the algorithm is - how does it handle specimen tilts around 90 deg?)
It'll also give you an idea of how accurately you need to know the actual tilt angles. Again, for low tilts, you'll probably need to know to within a degree or so but I'd bet that at higher tilts (45 to 60 deg) you may get away with knowing to the nearest 10 deg. If you do need to know the specimen surface tilt angles 'accurately', then I guess some sort of stereo pair procedure will give it to you?
I guess I'd be more worried about excitation volumes and X-ray paths to the detector that exit the sides of the cubes rather than the 'top' surface. This could be a problem irrespective of the tilt of the cubes, although you'll minimise it by rotating the specimen so that the surface you are analysisng is facing towards the detector and minimising the HT (to reduce excitation volume). It will also depend on the composition - if it's mainly low z, then the problem would be worse. Does the size of these cubes vary a lot and is it sufficient to check how apparent composition varies with size? If so, you might be able to work out a statistical extrapolation to bulk composition, although you'll need to analyse a lot of 'randomly' oriented cubes to get any sort of accuracy.
More practically, how much does the composition appear to vary and how accurately do you really need to know it? For example, if the apparent variation is within the error limits of the analysis anyway, then your problem dissappears! Alternatively, if you analyses a fairly large number of cubes (which you should be doing anyway, just like everbody else, to get good statistics .......:)...) can you demonstrate that there are clearly 2 or 3 seperate populations? If so, this might 'solve' your real pratical problem without even getting into the complexities of trying to do an accurate analysis.
Xianying Burany wrote: } } Good afternoon. } } I am looking for the phone number of Buehler Ltd. Please reply me if you } know. } } Thank you in advance. } } Sandy } 94765-at-udel.edu
1-800-Buehler 1-800-283-4537 and if you are looking for a competitor- 1-888-Struers 1-888-787-8377
--
Naresh Shah Research Associate Professor, University of Kentucky Consortium for Fossil Fuel Liquefaction Science (CFFLS) and Department of Chemical and Materials Engineering (CME) 533 South Limestone Street, Room 111 Lexington, KY 40506-0043 Phone: (606) 257-5119; FAX: (606) 257-7215; e-mail: naresh-at-pop.uky.edu
Beat Frey wrote: {I am wondering whether any Listserv subscribers have any experience {with the new PULSSTAR preferably in biological applications {that might help us guide our decision.
Dear Beat,
We are a materials science laboratory - but anyway: We have had very good experience with the PULSTAR digital pulse processor. The increased count rate is very useful - in particular when you are doing linescans or X-ray mappings.
If you are working at high count rates you will encounter some deterioration of the energy resolution; but in the cases where this is a problem, you still have the possibility of choosing a low count rate with the normal energy resolution.
Best wishes, Joergen
-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--at--at--at- J. B. Bilde-Soerensen Materials Department Risoe National Laboratory DK-4000 Roskilde Denmark
Can someone please point me to make/model/contact info for scanners that will digitize directly standard glass microscope slides that contain histological sections? Thanks so much.
I'm trying to get an antibody to work on glut fixed, osmicated, epon blocks. I have a very high background of the secondary binding to the sections on the copper grids, and cannot discern if the antibody is working at all, beyond the background "noise". Has anyone been successful with this, or am I wasting my time by trying to get this to work. For my purposes, I cannot use lowicryl, or non-osmicated tissue. Any suggestions?
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Thank you for your responses, but I fear I may have posed the wrong question. I am looking for oxide on the surface in more of an image analysis point-of-view, than quantifying the thickness of the oxide. I am considering the oxide to be like paint, and want to find the percentage of the wire's surface uncovered by the "paint". Typically, we have taken several photographs of the surface and crudely calculate the surface covered in oxide.
I hope I've explained myself a little better this time.
Marc C. Brande, MS, Founder wrote: } } Can someone please point me to make/model/contact info for scanners } that will digitize directly standard glass microscope slides that } contain histological sections? Thanks so much. } } Marc
I understand Polaroid makes one called Path Enabler. I have no personal experience with the product. -- Richard Miller M.D. Hematopathology "alles ist nur ein ubergang"
Does anybody know of accepted procedures to quatitatively evaluate immunehistochemical stainings. I am especially interested in evaluation methods for enzyme driven detection which has the problem of possible nonlinear relationships between signal and antigen abundance. Are there references to this problem?
Christian -- ************************************************************ Christian Broesamle email: broesam-at-hifo.unizh.ch Brain Research Institute tel: +41 1 385 6333 University of Zurich fax: +41 1 422 2262 CH-8029 Zurich http://www.hifo.unizh.ch Switzerland ************************************************************
Wonder if you have (and what kind) an EDS system? One with a light element detector (for O)? Some EDS sys will let you compare spectra/search a list for best fit. If you can obtain (standards) references of metal with various oxide film thicknesses, you could (using identical acquisition parameters) then compare the unknown spectrum to a series of references for the best fit. Once standards are collected, this should be quick and easy.
Another possibility is quantitative BSE. This would require similar standards. Digital image collection and manipulation would help quite a bit. Comparing the BSE coefficients (brightness at same op. parameters) to the standards will let you pick the closest film thickness. This method, depending on the oxide thickness, may require the use of 10 kV, 5kV, or less.
In either case the specimens will have to be "squeaky" clean to avoid false indications.
Have Fun!... Woody White Babcock & Wilcox Research Also -at-: woody.white-at-worldnet.att.net http://www.geocities.com/capecanaveral/3722 ______________________________ Reply Separator _________________________________
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Hello,
Often I am required to determine how clean a sample of strip steel is by comparing the amount of oxide remaining on the surface to the area scanned on SEM. The method we currently use is extremely time-consuming and uses up many polaroid films. Does anyone know of a simple way of finding the ratio of oxide left on a surface to bare metal? Is there any software available that may be of help?
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Hi chris. Sorry for the delay in getting you this. As we spoke about before I will need the Photoshop 4.0 upgrade for the PC. I don't know if it is relevant or not but I am running win95. The serial # you requested is"PWW300B1126201-337" Additionally I need: Maxtor 2.0 G hard drive , catalog #DR4875, price $269.95.
Epson stylus color inkjet, #PR11427, price $379.95. If you sell the ink, please send me a backup of each. I would appreciate if you could e-mail me back the total as well. This order was taken out of the MicroWarehouse catalog volume 40.0 so let me know if there are any price differences.
Customer # 6646947 PO# L02034
Ship to: Scott Whittaker University of Florida 218 Carr Hall Gainesville, FL 32653
Let me know if there is anything else you need and thanks.
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I believe that this forum is also very good source of information on this topic if you could be a little more specific. Many of the subscribers to this list have direct experience imaging polymer fibers, and could perhaps help you with your questions.
Regards,
-Bob ******************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com ********************************
I had a question concerning the imaging of coated and non-coated polymer fibers in scanning electron microscopy. If you could give me any useful websites and/or journal articles that are related to this topic it would be greatly appreciated. Susan Lloyd salloyd-at-eos.ncsu.edu
I have a microchannel plate on my 6300F JEOL. It performs extremely well in both modes - compositional (A+B) and topographical (A-B), and in low KV applications. It was purchased in 1995. If purchasing a new detector it is well worth evaluating this type, I'm sure there is more than one manufacturer. The one I have was built by Galileo Electro-Optics Corp. in Mass. (800-648-1800). No interest in the company.....just providing info.
********************************************************** Jake Schaper Product Analysis Lab Advanced Digital Consumer Division Motorola, Inc. 1300 N. Alma School Rd. Chandler, Arizona 85224 Mail Drop CH240 Phone 602-814-4756 **********************************************************
--------------------------------------
I'm sorry to join the discussion late, but I haven't checked my email for about a week.
I didn't see any replies recommending the micro-chanell plate as a solution to the low kEv situation. My last impression of these systems was that they were state of the art. They work very much like a photo-multiplier, and provide the high gain and very good signal to noise ratio.
If one were seeking a BSE system for low accellerating voltage situations, it would be worth contacting GW electronics in Norcross, GA. I have no interest in GW, other than seeing that the best technologies be evaluated.
One comment about the micro-chanell: the last time I looked, they didn't offer a topographic contrast mode (neither does the robinson). Why doesn't someone build a micro-chanell plate with 2 or 4 separate detector elements and high voltage supplies so the signals can be fed into a traditional amplifier which offers atomic number and topographic contrast? Sort of a multi-detector version of the micro chanell. Is there a practical reason this can't be done, or is it a commercial reason?
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Slovenian Society for Electron Microscopy Austrian Society for Electron Microscopy Croatian Society for Electron Microscopy Czechoslovak Society for Electron Microscopy Microscopy Society of Hungaria Italian Society for Electron Microscopy _ _ _ _ _ _ _ _ _ _ _ _ _
DATE AND LOCATION
MCEM 97 will be held from Sunday 5 to Wednesday 8 October, 1997 at Grand Hotel Emona Convention Centre, Portoroz, Slovenia (EU).
SCOPE OF THE CONGRESS
The aim of the Congress, which is held under the auspices of CESEM, is to allow the scientist to exchange the most recent results and technical advances in the development and application of electron microscopy.
The main topics are:
Ultrastructural localization of molecules Cryotechniques EM in cell structure and function EM in pathology EM of microorganisms New methods in analytical microscopy EM in material science Surface analysis Image analysis Digital processing and stereology Instrumentation and tools
SCIENTIFIC PROGRAM
The program will consist of invited lectures, oral presentations, poster presentations, and round table discussions. A limited number of abstracts will be selected for oral presentations.
ABSTRACTS
All those intending to participate at the Congress are welcome to submit an abstract. Deadline for abstract submission is April 30, 1997. Details for the format of the abstracts will be given in the Second circular (January 1997).
TIME SCHEDULE
January 1997: Second Circular and call for papers April 1997: Deadline for abstracts July 1997: Deadline for registration
CONGRESS LANGUAGE
The official language of the Congress will be English.
EXHIBITION
Ample space, immediately adjacent to the lecture and poster rooms, will be available for the exhibition of equipment, leaflets and books. Interested companies should contact the Congress Secretariat.
ADDRESS OF THE CONGRESS SECRETARIAT:
MCEM'97 Ceramics Department "Jozef Stefan" Institute Jamova 39, 1000 Ljubljana Slovenia
} I'm trying to get an antibody to work on glut fixed, osmicated, epon } blocks. I have a very high background of the secondary binding to the } sections on the copper grids, and cannot discern if the antibody is } working at all, beyond the background "noise".
First and foremost: don't use copper grids; use nickel or gold. Only some antibodies will still react in osmicated Epon sections - I presume you know yours will or at least have evidence that it survives tough treatment. There are standard methods to use: eg Ross A: Postembedding Ultrastructural Immunocytochemistry of Neural Antigens on Tissues Previously Processed for Routine L and EM....J EM Technique 5:81-90 (1987). Lastly, the smaller the gold particle, the more sensitive the reaction.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
I concur with the previous posting.... The effect on the quant with tilt will depend on the degree of tilt, the matrix, and the energy of the x-ray of interest. Using the lowest beam potential and highest x-ray (line) energy practical will minimize adsorption losses/corrections.
Suggestion, if you can prepare a polished specimen of similar composition... Using the polished material and the appropriate SEM conditions, collect spectra at several tilt angles. Collect a similar spectrum from the cubic specimen. The shape of the unknown (specimen) background may then be matched to the reference backgrounds and the associated tilt used for your calculations.
I have a question about standardless semi-quant work that I am doing, involving take off angle. ...SNIP... I want to examine a powder that has cubic structures within it. ...SNIP... of its position in space? I think the answer is yes and I've got a problem. My problem is, how do I perform standardless semi-quant work on the cubes within my powder, to examine homogeneity within the sample from one cube to the next?
Thank You in advance. Mark Darus General Electric Materials Characterization Laboratory Darus-at-cle.dnet.ge.com
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Hi! Need advice on staining human neutrophils for cytoskeletal proteins. My protocol works fine for other blood cells, but not for neutrophils. After fixing in 2% paraformaldehyde and permeabilization with 0.2-0.5% Triton there is no staining even for actin(Rh-phalloidin). I tried to use gluteraldehyde for fixing, but I get huge autoflourescense. Anya Shcherbina (shcherbina-at-cbr.med.harvard.edu)
Message-Id: {9610291825.AA22299-at-MIT.MIT.EDU} X-Sender: tonygr-at-po9.mit.edu X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
This is in response to Beat Frey's posting about the Pulstar pulse processor.
Beat does not say whether the application is to SEM or TEM. However, it is important to understand what EDX maufacturers are trying to do in developing their products.
The quality of the final analysis made with EDX depends upon the number of x-rays detected, and the signal to background ratio in the spectrum. The number of x-rays detected depends, of course, upon electron beam current and the time for the analysis, but also on the solid angle the detector subtends at the sample, and the ability of the system to process (i.e. measure) the x-rays. The signal to background ratio is partly determined by physics, but is improved by high energy resolution in the detector and analyzer system. Each x-ray takes a finite time for analysis; during that time, if a second x-ray arrives, it is lost (and, in some conditions, the first x-ray is also lost). This leads to the familiar "Dead time" which is reported, in one way or another, by all EDX systems. A pulse processor which processes x-rays faster leads to a lower dead time, and therefore less loss of x-rays. However, this is only useful if the loss of x-rays was originally significant. If the dead time is below about 25% the loss of x-rays is quite small.
While I'm not familiar with Noran's current product line, I would assume from Beat's comments that the Pulstar is their newest pulse processor, which probably is capable of a throughput of many tens of thousands of counts per second. However, it is important to understand that this specification assumes the x-rays will reach the detector in the first place. This leads back to my opening comment. Are the experiments capable of generating so many counts? If the microscope is a TEM, the samples are biological thin sections, and the probe size small, then perhaps not. This is not to say that upgrading to the new pulse processor would not be worthwhile otherwise, for it would undoubtedly also give you better energy resolution at lower count rates.
The fundamental question to be answered is the following: Do you have to adjust your experimental conditions to get the dead time on your current system down below 50%? (or, to rephrase, would you be able to use a larger beam current, but at the moment you don't because the x-ray count rate would be too high?) If the answer to either question is "yes", then the new pulse processor would benefit you. It will also give improved energy resolution, and will undoubtedly be serviced by the vendor for longer into the future.
My comments apply to all EDX systems, regardless of manufacturer; there may well be other features specific to the Noran line that would be benefits to you, too.
Hope this helps,
Tony Garratt-Reed. ********************************* ** ** ** Anthony J. Garratt-Reed ** ** MIT Room 13-1027 ** ** 77 Massachusetts Avenue ** ** Cambridge, MA 02139-4307 ** ** USA ** ** ** ** Ph: 617-253-4622 ** ** Fax: 617-258-6478 ** ** ** ********************************* *********************************
We offer the SprintScan 35 that will scan 35mm slides and negatives. There is an optional "Path Scan Enabler". This option acts as a carrier for a glass microscope slide to be scanned by the Sprint Scan 35.
There are three models available, 10 bit/1950 dpi, 10 bit/2700 dpi, and 12 bit/2700 dpi. They operate on both PC and Mac platforms.
The scanning software provides for a preview mode that offers image optimization prior to the actual scan.
Please contact me if you require additional information or visit our website at www.polaroid.com.
John D. Warren Southern Sales Manager "see what develops" Digital Photographic Imaging Group Polaroid Corporation
4525 Leonard Parkway Richmond, Virginia 23221-1809 Office 804.254.1011 Fax 804.254.1013 Internet warrenj1-at-polaroid.com
Can someone please point me to make/model/contact info for scanners that will digitize directly standard glass microscope slides that contain histological sections? Thanks so much.
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Regalado" {eregalad-at-cajeme.cifus.uson.mx}
THIS IS THE LMDC EQUIPMENT LIST. Tuesday 29 October 1996 Here you will find our SPECIALS which we wish to offer, NEW ITEMS when they arrive, and DISCOUNTS when we run them. If you wish to be on this list, please send return empty mail with "REMOVE EQUIPMENT LIST" in the header. Happy and money saving shopping. If you pass along our address to others with similar purchasing interests, we will appreciate it !
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Rachel: While immunocytochemistry on osmicated tissues sometimes works, the reactivity is usually less than on non-osmicated tissues; we have also had some problems with background. I would suggest trying the following: 1. Avoid copper grids as sometimes there are reactions with reagents that can leave a precipitate on the sections. 2. Try to oxidize the osmium before incubation with saturated sodium metaperiodate or 10% hydrogen peroxide. 3. Try changing your blocking solution; if you are using BSA, maybe try ovalbumin, instant milk, fish gelatin or normal serum, or a combination. Sometimes adding Tween 20 (0.05-0.1%) will help. 4. Decrease the concentration of the primary and increase the incubation time, e.g. overnight or longer at 4C. You might also try using tris buffer at pH 8.2. 5. You might also increase the dilution of your secondary.
Good luck! Art Hand UConn Health Center ------------------------------------------------------------------------------ FORWARDED FROM: Hand,Arthur Received: from Sparc5.Microscopy.Com by msgate.uchc.edu (PostalUnion/SMTP(tm) v2.1.8d for Windows NT(tm)) id AA-1996Oct29.175940.1274.219612; Tue, 29 Oct 1996 17:59:40 -0500 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id JAA03270 for dist-Microscopy; Tue, 29 Oct 1996 09:35:48 -0600 Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.4]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id JAA03267 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 29 Oct 1996 09:35:46 -0600 Received: (from teitelba-at-localhost) by post.aecom.yu.edu (8.7.6/8.7.3) id KAA02692; Tue, 29 Oct 1996 10:44:42 -0500 (EST)
I'm trying to get an antibody to work on glut fixed, osmicated, epon blocks. I have a very high background of the secondary binding to the sections on the copper grids, and cannot discern if the antibody is working at all, beyond the background "noise". Has anyone been successful with this, or am I wasting my time by trying to get this to work. For my purposes, I cannot use lowicryl, or non-osmicated tissue. Any suggestions?
Again, a little more information would be helpful, but I can think of a few image analysis (IA) approaches if you have not already considered them. Sounds to me like you are on the right track though, at one time we went down the road of manual grid-counting until we zoomed into the '90's and splurged on IA software.
1) If the oxide is fully distinguishable from the unoxidized surface by some topographical means or contrast difference using light microscopy or SEM, there are a number of IA programs that could simplify this assay. We have performed similar analyses using a PC version of program called "Imagemeasure", version 4.02, by Phoenix Technology, Inc. This software is sold in modules (counting & sizing, morphometry, densitometry, etc.) depending upon your needs. We used the counting and sizing module to determine the percentage of damage to corneal endothelial cell surfaces when touched by a plastic implant.
2) Other responses have suggested using backscatter detection, and this is another possibility. Obviously, in this procedure the unoxidized surfaces will appear brighter and can be quantitated using an IA program similar to that mentioned above.
Disclaimer: No interest in Phoenix Technology, commercial or otherwise, but if you would like their address or phone # contact me directly.
Good luck with your assay; hope this is useful.
Regards,
-Bob ************************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com **************************************
Thank you for your responses, but I fear I may have posed the wrong question. I am looking for oxide on the surface in more of an image analysis point-of-view, than quantifying the thickness of the oxide. I am considering the oxide to be like paint, and want to find the percentage of the wire's surface uncovered by the "paint". Typically, we have taken several photographs of the surface and crudely calculate the surface covered in oxide.
I hope I've explained myself a little better this time.
"Marc C. Brande; MS; Founder" {mcbrande-at-sierra.net} Cc: HistoNet-at-pathology.swmed.edu, microscopy-at-Sparc5.Microscopy.Com, nih-image-at-soils.umn.edu
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Hi Marc,
I use the polaroid SprintScan 35 when I am scanning histological sections. I use with it the PathScan enabler. The SprintScan can be used with either Windows or Macintosh with the appropiate software and SCSI. You can get the Scanner at a computer store. The PathScan enabler is available from Meyer Instruments (713)579-0342, Houston Texas.
The scanner gives pretty good resolution from the entire slide. The files tend to be quite large. So you want to make sure you have enough RAM and space to store these files.
Can someone please point me to make/model/contact info for scanners that will digitize directly standard glass microscope slides that contain histological sections? Thanks so much.
(IMA Internet Exchange 2.03 (Beta 5) Enterprise) id 00012B79; Tue, 29 Oct 96 10:11:07 -0600 Received: from pathology.swmed.edu (pathology.swmed.edu) by SWAX32.SWMED.EDU (PMDF V5.0-7 #12086) id {01IB7MINIFNC9EDCMK-at-SWAX32.SWMED.EDU} for Ray_Ortiz_at_RLT011-at-ccmailgw.mcgawpark.baxter.com; Tue, 29 Oct 1996 10:12:07 -0600 (CST) Received: from 129.112.18.39 by pathology.swmed.edu with SMTP (Apple Internet Mail Server 1.1.1); Tue, 29 Oct 1996 10:15:25 -0600 Received: from diamond.sierra.net (207.135.224.253) by pathology.swmed.edu with
You will have received an Email "sales" bulletin from EM-at-MediaCity.Com. Please note that this individual has been removed from the listserver as having violated our rules despite previous warnings.
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All hate/flame mail should be directed to EM-at-MEDIACITY.COM, and office-at-lasermotion.com they have already gotten mine!
It's been a while since I did immunohistochemistry at the EM level, but when I was doing it I had a problem with high background stain that I was able to reduce by using nickel instead of copper grids. It had something to do with an interaction between the copper and the colloidal gold I think... I have never tried osmium fixed tissue, however, so I can't help with that part.
Karen
On Tue, 29 Oct 1996, Rachel Teitelbaum wrote:
} } I'm trying to get an antibody to work on glut fixed, osmicated, epon } blocks. I have a very high background of the secondary binding to the } sections on the copper grids, and cannot discern if the antibody is } working at all, beyond the background "noise". Has anyone been } successful with this, or am I wasting my time by trying to get this to } work. For my purposes, I cannot use lowicryl, or non-osmicated tissue. } Any suggestions? } } Thanks, } Rachel }
I'm putting together a course in metallography and failure analysis for a mixed senior undergraduate / graduate group. I would like to raise the intellectual content of the course beyond the typical technique survey and review of case studies and add a significant component of materials reliability and electrical/electronic FA. Any thoughts or suggestions would be appreciated.
Does anyone share my concerns about intellectual content of FA-type courses or have a positive example (outside of a short-course)? Several universities seem to combine FA with design mechanics to add some analytical content, but I would prefer to stay closer to materials failures while broadening the industrial scope. The incorporation of tribological failures (e.g. magnetic media) and electronic failures (electromigration, dielectric breakdown) seems to be woefully lacking in texts on the matter.
Thanks Daniel L. Callahan Assistant Professor Mechanical Engineering and Materials Science Rice University
We work with ethanol production by alcoholic fermentation in large industrial plants. One of the main controls is the microscopic examination of the yeasts in the several stages of the process. In order to improve this control we would like very much to develop or to buy a system to do the examination automatically. Is there such a system ? I mean, a system that resembles a Flow Injection Analysis, i.e. it takes a sample from the fermenter, dilute it, mix with the contrast inject it at the microscope slide (continuously?) take a sample image and analyse it. Any hint about it, mainly a contact person, would be very, very nice. I thank you in anticipation for your kind attention to our problems.
Best regards
Jaime Finguerut(Mr., chem.eng.) Cx.Postal 162 Piracicaba Sao Paulo BRAZIL 13400-970 fax 0055 194 29 8388
To everyone who gave me tips on removing epon blocks from their molds - I want to thank you. I knew microscopists were creative but I didn't realise just HOW creative everyone was!
This reply comes from the US distributor of ImageSlave. His address is jhilton-at-rmi.net.
} Dear MicroPeople: Reply to ImageSlave under NT. } } It will not work, since the HAL spec. is not met, the card is not PCi, the } software is not 32bit. However, it might work in NT using the DOS engine. } But what's the point? } } Jim Hilton } } } Jim, this just came through the microscopy listserv. Thought I'd pass it } along, in case you don't subscribe. } } } } } Date: Tue, 29 Oct 1996 09:06:43 +0800 } } } From: SCM!ATitkov-at-scmaust.attmail.com (SCM!ATitkov) } } } Subject: ImageSlave } } } To: Microscopy-at-Sparc5.Microscopy.Com } } } Mime-Version: 1.0 } } } Status: } } } } } } Dear Microscopists, } } } } } } Has anyone tried to use ImageSlave under Windows NT? } } } } } } Alexander Titkov } } } SCM Chemicals Ltd. } } } PO Box 245 } } } Bunbury WA 6231 } } } Australia } } } Ph (097) 808 505
Another reference that may be of interest is: Bendayan M, Zollinger M, 1983, "Ultrastructural localization of antigenic sites in osmium-fixed tissues applying the protein A-gold technique", J Histochem Cytochem 31:101-109.
A. Kent Christensen University of Michigan {akc-at-umich.edu}
---------------------------------
On Wed, 30 Oct 1996, Diana van Driel wrote:
} } I'm trying to get an antibody to work on glut fixed, osmicated, epon } } blocks. I have a very high background of the secondary binding to the } } sections on the copper grids, and cannot discern if the antibody is } } working at all, beyond the background "noise". } } First and foremost: don't use copper grids; use nickel or gold. Only some } antibodies will still react in osmicated Epon sections - I presume you know } yours will or at least have evidence that it survives tough treatment. } There are standard methods to use: eg Ross A: Postembedding Ultrastructural } Immunocytochemistry of Neural Antigens on Tissues Previously Processed for } Routine L and EM....J EM Technique 5:81-90 (1987). Lastly, the smaller the } gold particle, the more sensitive the reaction. } } } Diana van Driel } Dept Ophthalmology } Sydney University C09 } AUSTRALIA 2006 } } }
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Hi! Need advice on staining human neutrophils for cytoskeletal proteins. My protocol works fine for other blood cells, but not for neutrophils. After fixing in 2% paraformaldehyde and permeabilization with 0.2-0.5% Triton there is no staining even for actin(Rh-phalloidin). I tried to use gluteraldehyde for fixing, but I get huge autofluorescense. Anya Shcherbina
Message-Id: {199610301516.KAA12374-at-ns1.axs2000.net} To: MICROSCOPY BB {Microscopy-at-Sparc5.Microscopy.Com}
} I will be doing ultramicrotomy of paper which has been coated. Could I } please have suggestions and comments on an embedment process. } } Thank you } } Leah L. Dobbs }
Coated paper can be tricky. We've used Spurr's resin successfully - make sure the edges of the sample are clean (ie cut with fresh razor blade or scalpel) since these edges will be the route for the resin to pentrate. Long infiltration times, use of a rotator, and alternating vacuum to atmosphere cycling can also help (but avoid excessive time under vacuum and use appropriate safety measures).
There are some references available in the pulp and paper industry literature. You might want to search out articles by Dale Quackenbush - he used epon812 for embedding and cut high quality 2 micron thick sections with a sledge microtome! Most of his work is found in TAPPI magazine or The Microscope.
I hope this is a good starting point.
James Drummond Pulp and Paper Research Institute of Canada 3800 Wesbrook Mall, Vancouver B.C., V6S 2L9 Phone: (604)222-3200 Fax: (604)222-3207
I never use anything but Ni-grids -when I in the beginning happened to incubate the primary ab overnight and used coppergrids, the drop turned blue, so something definitely goes wrong there...
Like everybody else already said, there are antibodies that work with osmicated tissue, I have never tried it though. What I would like to add, is that I a couple of years ago I saw a paper from a japanese (?) group, that had reduced their osmium with potassiumferrocyanide, and that made their ICC work better. Maybe you can find that article? (I didn't keep it.)
} Thank you for your responses, but I fear I may have posed the wrong } question. I am looking for oxide on the surface in more of an image } analysis point-of-view, than quantifying the thickness of the oxide. } I am considering the oxide to be like paint, and want to find the } percentage of the wire's surface uncovered by the "paint". Typically, } we have taken several photographs of the surface and crudely calculate } the surface covered in oxide. } } I hope I've explained myself a little better this time. } } Thanks in advance, } } Tania Jones } } tania-at-dynamotive.com
Any 'automated' approach will rely on having something in the image that clearly distinguishes the oxide from the rest of the specimen. If you have that then most image analysis programmes will handle the problem within seconds.
If you have a 'modern' SEM, you can probably transfer the image directly into your image analysis computer. With older SEMs, you'll either have to connect your computer to the SEM via a frame grabber, have an image scanned.
As to regards software, I'd suggest starting with free/shareware - NIH Image is a good option. This will give you a chance to evaluate the procedure without committing too much money.
Mr-Received: by mta SRVR05.MUAS; Relayed; Thu, 31 Oct 1996 09:50:16 -0500 Mr-Received: by mta SRVR05; Relayed; Thu, 31 Oct 1996 09:50:16 -0500 Mr-Received: by mta SRVR01; Relayed; Thu, 31 Oct 1996 09:50:18 -0500 Disclose-Recipients: prohibited MSA MICROSCOPY MAILING LIST {MICROSCOPY-at-Sparc5.Microscopy.Com} Message-Id: {4016500931101996/A20861/SRVR05/11AAFA720F00*-at-MHS} Autoforwarded: false Mime-Version: 1.0 Content-Type: TEXT/PLAIN; CHARSET=US-ASCII Content-Transfer-Encoding: 7BIT Importance: normal Priority: normal Sensitivity: Company-Confidential Ua-Content-Id: 11AAFA720F00 X400-Mts-Identifier: [;4016500931101996/A20861/SRVR05] Hop-Count: 2
One paper is entitled, "Improved Method for Post-embedding Cytochemistry Using Reduced Osmium and LR White Resin", H. Tamaki and S. Yamashina, J. Histochemistry and Cytochemistry, 42:1285-1293, 1994.
Best regards, Walt Bobrowski Parke-Davis Research
} I never use anything but Ni-grids -when I in the beginning happened to } incubate the primary ab overnight and used coppergrids, the drop turned } blue, so something definitely goes wrong there... } } Like everybody else already said, there are antibodies that work with } osmicated tissue, I have never tried it though. What I would like to add, is } that I a couple of years ago I saw a paper from a japanese (?) group, that } had reduced their osmium with potassiumferrocyanide, and that made their ICC } work better. } Maybe you can find that article? (I didn't keep it.) } } Margareta
Not knowing exactly what yeast cell parameters your interested in it would seem to me that an intergration of a flowcytometery system and a machine vision / auto-recognition system might be a way of acheiving what you need. Since you are involved in industrial fermentation a bigger concern would be the wide location of fermentation vats and an analysis system. (.... allowing my imagingation to run wild: the possibility of floor track guided automatic sample withdrawal system collecting and transfering samples from vat to analysis workstation....mmm)
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
For anyone who has cut their fingers using beem molds and razor blades there is a new development called "easy molds". The epoxy bullets and samples remove easily so a razor blade is not necessary to remove the samples. The person to contact regarding these new molds is:
Message-Id: {9611010108.AA1308-at-pho903.sbphrd.com} To: microscopy {microscopy-at-Sparc5.Microscopy.Com} {Beverly_E_Maleeff-at-sbphrd.com}
PHILADELPHIA SOCIETY FOR MICROSCOPY
NOVEMBER 1996 MEETING NOTICE
Wednesday, November 13, 1996
New Frontiers in Thoracic Imaging
Warren B. Gefter, M.D. Department of Radiology University of Pennsylvania Medical Center Philadelphia, PA
This presentation traces the rapid evolution of the following exciting advances currently taking place in thoracic imaging: digital (computed) radiography, progress in computed tomography (CT) including high-resolution CT, helical and electron beam (ultrafast) CT; magnetic resonance imaging (MRI) and volumetric image display methods such as virtual endoscopy. These methods are providing multidimensional displays of cardiopulmonary anatomy and physiology. Examples of the imaging provided by these techniques will be shown. In addition, ongoing developments in volumetric image processing techniques are providing methods to display these complex multidimensional datasets, useful in both the clinical and research realms.
DATE: Wednesday, November 13, 1996
PLACE: Laboratory for the Research of Science and Materials (LRSM) Building, 33rd and Walnut Street (map enclosed). Parking is available behind the LRSM Building after 5:00 PM.
TIME: 5:30 PM Social hour, hosted by our meeting sponsors
6:30 PM Dinner Members $12.00 Student members $6.00 Non-members $15.00
Menu: Beer, wine, assorted soda and bottled water Munchies
Baked ziti with ricotta, mozzarella and parmagiana cheeses Breast of chicken House salad with Italian dressing Chef's vegetable du jour Rolls and butter
Golden cake with chocolate icing
Coffee, decaf or tea
7:30 PM Speaker
Reservations will be taken by Ms. Pat Overend at the University of Pennsylvania, 215/898-8337. Deadline for reservations will be Monday, November 11. If you have any questions regarding the meeting please feel free to contact Rollin Lakis of the Executive Council. Cancellations must be received by Ms. Overend no later than 5:00 PM, November 11, 1996.
RESERVATIONS ARE REQUIRED FOR DINNER. We cannot guarantee you a meal if you do not make a reservation by the above deadline.
Message-Id: {2.2.32.19961031204014.0069e4cc-at-darkwing.uoregon.edu} X-Sender: mshaf-at-darkwing.uoregon.edu X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 02:27 PM 10/31/96 +0000, Marc wrote: } } Would someone kindly explain how to convert dpi (dots-per-inch) to } pixels? } } I would like to evaluate scanner resolution (ie. 2700 dpi) in terms of } pixels. } "Dots" (as in dots per inch) are usually equated to pixels ... that is, if you scan a 35mm slide (1.25"" by 1") at 2700dpi you'll end up with approximately a 3400 by 2700 pixel image. If this is then printed with a 300dpi printer then its final size would be (approximately) 11" by 9".
The confusion generally sets in when some printers demand you are knowledgeable of what it is capable of printing in terms of "lines per inch" (lpi), which has much to do with its inability of printing one of the 16M colors as one "dot" (dye-sub printers have this ability), but instead need a small matrix of dots to dither CMYK into a resemblance of one of 16M colors. Thus, if a 300dpi printer (ink jets, color laser) needs an 8by8 matrix to create one of these colors then its true capability is 300/8 or ~37 lines per inch.
So much for the math ... there is a lot of gray area here such as "apparent" capabilities for printers to do better than this in spite of the math ... so begin with an understanding of the terms and experiment ...
hope this helps ...
cheers, shaf {/} \ {/} \ {/} \ {/} \ {/} \ {/} cognito, ergo zZOooOM {/} \ {/} \ {/} \ {/} \ {/} \ {/} Michael Shaffer - mshaf-at-oregon.uoregon.edu - mshaf-at-darkwing.uoregon.edu Electron Microprobe Facility - Geological Sciences - University of Oregon http://darkwing.uoregon.edu/~mshaf/shafhome/