Although X-ray spectra can do this sort of work, isn't it true that for the most part the electron spectroscopies tend to do the job easier? I'm thinking here of mainly of ESCA/XPS (i.e. the photoelectron spectroscopies).
Once you have collected all the wisdom of the world on Screens please post a summary here for all to read.
The same should apply to everyone that posts a question and collects answers. Sometimes as Pierre pointed out not all replies go to the listserver and you would be "paying your dues" by providing a concise summary of all reponses for all subscribers to read and possibly file for future use.
Subject: Time:12:22 PM OFFICE MEMO Ion Tech Address Date:6/1/94 I have an address for Ion Tech Inc. at 2330 East Prospect, Ft. Collins, CO, 80525. Tel: 303-221-1807; Fax: 303-493-1439. My source gives offices in Germany and Japan, but none in the UK, so this may not be the organization you are interested in.
Is there fluorescent stain that can be applied as easily as DAPI or Hoescht stains that have excitations and emissions in the same wavelength as FITC or Rhodamine?
Thanks, David
=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= David L. Hirschberg bnhirsch-at-wicc.weizmann.ac.il Department of Neurobiology (972) (0)834-2127 (0)834-2412 work Weizmann Institute of Science (972) 847-4805 home Rehovot 76100 Israel (972) 834-4131 fax
} Although X-ray spectra can do this sort of work, isn't it true } that for the most part the electron spectroscopies tend to do } the job easier? I'm thinking here of mainly of ESCA/XPS (i.e. } the photoelectron spectroscopies). } } Nestor Zaluzec } ANL EMCenter } It depends on the spatial resolution you require. Here in Bristol we have a VG Escascope small area/imaging x-ray photoelectron spectrometer. The smallest area you can obtain XPS spectra from is 50um diameter, so that is the smallest area from which you can obtain chemical shift information (as long as you are prepared to wait a considerable time for the spectrum to appear out of the noise). In the imaging mode, {10um resolution can be obtained,where you image a chosen photoelectron energy, so the distribution of a particular chemical shift can be seen, at that sort of resolution. Newer instruments canimage down to 1 -2um. Nevertheless, *if* you can obtain the data you want on an sem or probe, then maybe, and I don't really have any experience of looking for chemical shift in wdx or edx spectra, you can more easily do so in small regionsof the sample, and perhaps with shorter acquisition times.
Keith Hallam Interface Analysis Centre University of Bristol England
Yesterday I sent a message about the Hitachi vs AMRAY problem. Unfortunately after sending it I read it again and it seemed as if I was giving a dim opinion of the scopes AMRAY produces. That was not my intention at all. I got the company mixed up with another company I had dealings with and with the other company I did have bad experiences with. It was another American company and I haven't heard anything about that company in a long time. It could be that company is no longer in business. I don't know. To AMRAY I humbly apologize. I know your sales people and have a deep respect for them. It was totaly unprofessional for me to make a mistake like this without reading what I sent out. Again, I humbly apologize.
There is a magnificent handbook of fluorescent probes/dyes published by Molecular Probes, Inc. of Eugene, OR.: FAX (503) 344-6504 or telephone (503) 465-8300. It serves as their catalog, and gives really extensive, comprehensive descriptions AND bibliogr aphies. They do have worldwide distributors, but I don't see any listed for Israel--let me know if I can help any further. Good luck. Grace Kennedy, Scripps Inst. of Oceanography/UCSD (neurobiology, too.)
Hello, humble Masters student here. My question is regarding the standardization of one samples x-ray spectrum to another. If my SEM was nice and stable I suppose I could just take a spectrum for a certain period of time. Or if I had some nice low noise amplifiers I could look at the sample current and integrate that. However, my SEM isn't very stable and I'm not sure I want to mess around with the sample current. Has anyone heard of any other methods or can point to some articles regarding this subject.
Thanks in advance. Dan Henne Simon Fraser University Vancouver, Canada. email: henne-at-sfu.ca
This is in response to the message regarding DAPI nucleic acid stain. The filter set needed for the DAPI fluorescent stain (bound to DNA) is the UV excitation filter set; EX max 359 nm and EM max 461.
I have a Hitachi SEM and TEM and service is good. The rep is very responsive to my scheduling problems and works hard to get the scope running quickly. There is no question about his competence. Neither scope has ever been down for more than a few days for lack of parts. Yet I have still not been totally satisfied with Hitachi service because they depend on the user to know when there is a problem. When Hitachi services a scope, they fix that problem and leave. They perform no other routine checks to catch problems early or to deal with problems that might go unrecognized by a microscopist, particularly if new to EMs. Further, they have no PM checklist to demonstrate to the microscopist that the scope really is running at peak performance. In short, the Minnesota rep is competent and very helpful but is not fully supported by the organization. AMRAY used to have a marginal reputation but their company seems to have undergone a complete transformation. I attended one of their user meetings and it was apparent that their discipline and attention to user needs was unmatched by other organizations I had examined. I spoke to several participants and they were extremely enthusiastic about AMRAY service and many of these people had prior experience with other big-name manufacturers. For example, after AMRAY services a scope, they do a routine check on all switches and other items that are reasonably easy and fast to check. AMRAY also uses an extensive check list on their PM which is presented to the microscopist along with test photos. I also had the impression that AMRAY reps are better trained before they are sent into the field solo. When my rep started here, he didn't know how to stigmate the TEM! Of the service meetings I attended (Hitachi, AMRAY, Cambridge, and JEOL), AMRAY was by far the most aggressive in extracting grievances from their users. The AMRAY people running the meeting worked their way down a rather long list, "Has anybody had trouble with response time? Has anyone had trouble getting parts." They spelled out their standards for response times so people knew what to expect. I was impressed. If this meeting (now about 2 years in the past) was typical of the organization, they must have good scopes and service. I must note that I have no personal experience on an AMRAY or with their service. My impressions result from this one AMRAY user meeting.
Are there any ultrastructural biologists in the email network that have used any techniques for demonstating acetyl cholinesterase receptors at the ultrastructural level ? Does anyone know of any source for peroxidase conjugated, gold conjugated alpha bungarotoxin or an antibody that has been raised to alpha bungarotoxin?. I would appreciate any information.
Terry Robertson
Dr Terry A. Robertson Telephone: 61-9-3462935 Department of Pathology Fax: 61-9-3462891 University of Western Australia Nedlands WA 6009 Australia Email: troberts-at-eosin.path.uwa.edu.au
After 2 years of debating about this I decided to start seeking a position out of the U.S.A. The areas I narrowed it down to are Costa Rica or Australia. I was wondering if anybody knew of any universities or private companies in these areas that might have need of an electron microscopist with 27 years experience. Actually, somewhere in Europe I would consider also, preferably Sweden, Germany or Switzerland. Thanks,
Does anyone know of a book that lists emission wavelengths for the elements in the 1000 to 2000 angstrom range? We can find some info where the wavelengths are listed by element, but we would like a listing of WLs in order of WLs, in order to check for interferences.
TIA,
Sue Smith National Steel Corp. smiths-at-mlc.lib.mi.us
Try and get in touch with Prof. Brian Gunning, Research School of Biological Sciences, Australian National University, P.O. Box 475, Canberra ACT 2601. He may able to suggest the names of persons who may know about openings for electron microscopists.
Can I please once again remind you that the listserver is not to be used by Job hunters looking for positions. I am sensitive to this issue as I have turned down many requests from both ANL personnel as well as outside users all qualified and many being personal friends, all who have ASKED to post this type of information and were turned down. Given that I just posted a notice reminding people of this only a week ago I am extremely disappointed in the recent postings. Once again, you are welcome to use the MSA/EMC BBS for these purposes, but NOT the Microscopy Listserver.
If you are not sure about a posting, I would be glad to have a quick look at it first before forwarding it to the listserver especially, if you are not sure whether or not it is appropriate. I have already done this many time for subscribers (most of which were by the way posted without modification). BUT, I donot want to convert this to a moderated, subscription only list as I really don't have the time. So please remember or refresh you memory on the "ground rules" you were sent when you first subscribed. I didn't ask for much, and your getting alot for almost nothing. I will also consider the suggestion that was sent to me by several subscribers of removing repeat "offenders" from the listserver, if someone gets abusive.
Dan Henne asked about standardizing beam current for taking x-ray spectra:
First of all I agree with what Bob Craig said in his nice n' concise summary of the do's and don't's of collecting "calibrated" x-ray spectra. I'll specifically address Dan's question of how to set up a calibrated, reproducible beam current with out using any kind of current meter. Of course, you won't know the *value* of the beam current, but at least you'll know it is the *same* value from sample to sample and day to day.
We do not have a specimen current meter nor a Faraday incident beam current meter, so what we do in my lab to standardize beam current is the following: Mount a small metal foil, like aluminum or copper, onto the edge of each stub that the samples are mounted on using carbon paint or double stick tape. Run a little collodal graphite around an edge or two to provide additional adhesive support and a conductive bridge to the foil. Then before taking spectra, move the electron beam over to the foil and look at the metal foil count rate. Then adjust the beam current using the condensor lens, or spot size, control to set an arbitrary rate, say 3,000 counts per second, near the high end of x-ray through-put for your system (especially for biological samples; may not need a high end setting for materials samples), but not more than, say, 33% deadtime. This value can be checked for stability periodically and should be checked for each new stub you put into the SEM. Thus you will be delivering the same incident beam current to your unknown samples. Of course, there is always some indeterminate error with any kind of instrumental setting, and the count rate fluctuates a bit from second to second due to detection and counting statistics in the electronics and the physics of beam current production. With our set-up, the fluctuation is no more than 100 counts per 3,000 and over a 100 second analysis time the fluctuations average out. Also, our SEM a Philips 500X, is quite stable. Provided the filament doesn't shift, the same 3,000 cps will be there hours later. We set the rate when we put a sample in, and check it about every 30 minutes to monitor any changes that may occur. If a change has occured, we must throw out some data and collect over again.
A few precautions: use only clean foils; actually image the foil surface at moderate magnification to make sure that you place the beam on a clean area without microscratches. Also, must set the stage tilt and detector-to-sample distance to some standard and reproducible setings, etc. as Bob Craig pointed out.
But if your SEM is as "unstable" as you make it sound, you will have to check the foil count rate immediately befor and after collecting a spectrum and keep only those spectra where the rate did not change during the analysis time. If it does change frequently, you may only be able to do limited relative quantitative analysis based on ratios.
One comment about the use of specimen current (if you're lucky enough to have a specimen current meter on your SEM) to set incident beam current. As Bob Craig said, you can't do it by monitoring the current on the actual specimen, UNLESS you pick the appropriate specimen; for example, keep a margin of the aluminum stub clean and free of specimen, paint, tape, scratches, etc, and measure the "specimen" or absorbed current there. This value is the total incident current minus the backscattered fraction, and this fraction should stay the same as long as you use the same metal target, kV, geometry, etc, etc, etc, etc. So you are sampling the same fraction of the total and by setting the same value of specimen current, you are then standardizing the incident curret. I would think that this would be another way, like my above mentioned indirect technique based on foil x-ray production, to consistently set a constant incident beam current.
--
Gib Ahlstrand 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
We are in the market for a backscatter detector for our Cambridge 180 SEM. I ask your opinions on makes,models, etc. for such an item. We mainly are a life science (pathology) lab thus any info relating to this area is appreciated. Also any info, opinions, etc. on multi-channel-plate detectors vs backscatter detector? Are they worth the extra dollars? (for a life science lab)
I was not sure wheather you are looking at plant cells or animal cells. If it is plant cells, there several publications. But if you want a single source perhaps the one by Behnke would be most helpful:
Behnke, H-D. (1991). Nondispersive protein bodies in sieve elements. A survey and review of their origin, distribution and taxonomic significance. Internatl. Assoc. Wood Anat. (IAWA) Bulletin 12:143-175.
We have been dealing with AMRAY for 17 years now. There service has been outstanding here in the Bay Area: they respond quickly, home in on the problem, and do what needs to be done. They do routine maintenance. Our 1000A microscope is getting along now but it has always made very good pictures and remains an excellent low-resolution instrument: it runs and runs with little down time.
Jacob Bastacky Jacob Bastacky, MD Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Message-Id: {9406040024.AA03054-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: XEDS;SEM;Standardizing spectrum Orig-Author: {Daniel Henne {henne-at-sfu.ca} }:ddn:wpafb ----------------------------------------------------------- Hello, humble Masters student here. My question is regarding the standardization of one samples x-ray spectrum to another. If my SEM was nice and stable I suppose I could just take a spectrum for a certain period of time. Or if I had some nice low noise amplifiers I could look at the sample current and integrate that. However, my SEM isn't very stable and I'm not sure I want to mess around with the sample current. Has anyone heard of any other methods or can point to some articles regarding this subject.
Thanks in advance. Dan Henne Simon Fraser University Vancouver, Canada. email: henne-at-sfu.ca
Ion Tech Ltd was taken over a few years ago - I cannot remember by who. However, they are still in business but trading under the name of Atom Tech Ltd.
Atom Tech Ltd can be reached in the UK at:
Island Farm Avenue, West Molesey, Surry KT8 2UZ, England.
I have been unsuccessful in locating ANY information on the International Symposium on the Structure and Properties of Dislocations in Semiconductors to be held in 1995 (I believe in Italy). Would anyone happen to have anything on this conference or direct me to someone who does? Any assistance would very much be appreciated. Thank you.
H. R. Kolar Center for Solid State Science Arizona State University FAX: (602) 965-9004
Apologies to anybody who received this twice due to cross posting.
Every once in a while somebody asks for a survey of user fees for multi-user facilities involving LM or EM.
The discussion of fees has been very informative.
We were wondering, in addition to fees, how these facilities are administrated.
For instance: o What is the administrative hierarchy? o What is the salary and rank of the director of day-to-day operations? o What are the responsibilities of this person or people? o Who contributes the equipment to the facility and how are these resources allocated to users?
Any comments would be appreciated.
Thank you.
Michael Cammer cammer-at-aecom.yu.edu cammer1-at-telico.bioc.aecom.yu.edu
I have a microscope that came with a camera adapter that, among other things, has a film holder for 2 3/4 x 3 1/2 sheet film. Has anyone ever used film of this size? I would certainly appreciate any information that I could gather. I am not familiar with this size, or why it would be used.
At the HVEM we use 6.5 * 9 cm film (~= 2 3/4 x 3 1/2). This film is satisfactory for our purposes and the larger standard size will not fit in the camera. Either Kodak 4489--a very fine grain film which is our usual choice-- or SO163 can be obtained in this size. We also have some old x-ray film cut to this size which is used primarily for recording diffraction data. If you de- cide you don't want to get into this size film, could you please send me a de- scription of the holders you have? By some miracle, they may be compatable with the AEI holders we use. Thanx.
The EMMPDL has a contributed MC program called MacSim. Here is the abstract...
Nestor
Title :MONTE-CARLO SIMULATION (Z's MC v. 1.1) Keywords :.Electron-beam, Monte-Carlo simulation, energy loss, backscattered electrons, Rutherford cross section, Mott cross section. Computer :.Macintosh Operating System :6.0 or 7.0 Programming Language :Pascal (Think Pascal) Hardware Requirements :standard Author(s) :.Zbigniew J. Radzimski. Correspondence Address :.Analytical Instrumentation Facility North Carolina State University Raleigh, NC 27695-7916 Abstract: The current program is the further development of the program written many years ago by R. Myklebust and D.C. Joy and modified later by J. C. Russ (J. of Computer Assisted Microscopy, 1990, 2(2), p. 59). It calculates various parameters related to electron-beam interactions with solids related to absorbed and backscatetred electrons. It accepts multi-element and multi-component structures. It uses Rutherford or Mott cross section for scattering. Special thanks to Zbigniew Czyzewski and David Joy for providing Mott tables (see J. Appl. Phys., 1990, 68(7), p. 3066).
I know someone who is helping to set up an EM lab. They are interested in finding out where and how they might send someone to get training. I know that the various makers of EMs offer courses on how to use their instruments, but where can they get training on processing, sectioning, and other techniques used for boilogical TEM. (used for examining ultrastructure). Any information will be helpful.
There are several universities that offer courses in the techniques of biological electron microscopy including Cornell. Some years ago, I think the Microscopy Society of America (MSA) sponsored a survey of EM courses offered in the U.S. universities. If you get in touch with MSA they may be able to send you a copy of the survey.
Partha M.V. Parthasarathy Section of Plant Biology Cornell University, Ithaca, NY 14853. USA Tel: 607-255-1734 Fax: 607-255-5407
Our facility is designed to be totally self funded. Administration heirarchy is:
Faculty director (25% time) Sets goals, policy, etc. Consults with users and helps design projects Sees to overall day to day administration
Facilities engineer (full time) Maintains microscopes (we 2 SEMs, TEM, IVEM) Trains users Developes film Helps with specialized procedures
2 EM technicians (full time, each) Processing and embedding Specialized procedures Immunocytochemistry Enzyme cytochemistry Autoradiography Etc. Sectioning Printing Maintain EM records Billing
Contact Kodak in Rochester, NY. Ask for the Scientific Advisor/Advice Department.
Sue Smith for Sam Purdy -at- National Steel, Trenton, MI
On Fri, 3 Jun 1994 ARGIL-at-delphi.com wrote:
} I have a microscope that came with a camera adapter that, among } other things, has a film holder for 2 3/4 x 3 1/2 sheet film. Has } anyone ever used film of this size? I would certainly appreciate any } information that I could gather. I am not familiar with this size, or } why it would be used. } } Thanks, } } Arthur Gillman } Princeton, NJ } }
Message-Id: {MAILQUEUE-101.940607084003.448-at-bunyip.ph.rmit.edu.au} To: microscopy-at-anlemc.msd.anl.gov
Re : Fees for use of facilities in Em Units
Since we have only recently joined this group, we missed the comments on user fees which have apparently been circulating from time to time. Does anyone have a summary ? We are interested in not only fees for individuals or groups external to the university, but also we are obliged to cross charge for all teaching and research within the university to support the Unit. What scale of fees are commonly used for SEM, TEM,EPMA, spec prep from simple to complex ? Are there "discounts" for local users or are there different rates ? Are the rates different for expert users ? etc, etc.... THank you in advance for the comments.
John Millar EM Unit, RMIT, Melbourne Australia jjmill-at-rmit.edu.au
There are at least 2 very good BSE detectors assemblies namely Robinson BSE and Oxford Instruments P/L TETRA BSE. Robinson BSE is well known for its performance including biological subject but I was most impressed by per formance of TETRA BSE. It is auto-biased with four independent quadrants, which can give variety of BSE signal combination and working with a electron beam treshold of 0.1 nanoamp--I think-- it is a jewel. Have seen it working at ElectroScan Corp- Boston installed with E-SEM. Regards and good luck with the purchase , Wis Jablonski CSL Uni Tas ,Tasmania, Australia
Greetings, These are follow-up questions for Jay Jerome's posting regarding his unit: how many people does your lab serve? What percentage are from "in-house" (your department/division) vs. those from "outside"? Do you charge different fees based on the affiliation of the user and their level of expertise? How do you control the "quality" of the user? I have worked in the Cornell TEM Facility run by M.V. Parthasarathy, which seems somewhat similar to yours (with the exception of no in-house technical support for the instruments), where a user could pay a flat fee for a certain period, plus extra costs for special equipment, e.g., high-pressure cryofixation. A user could also pay on a per hour, plus materials basis. Do you have such a scheme as well? My department, which has a simple facility with an SEM and a TEM, charges virtually no fees to department users, except for the materials (films, stuff used in processing material, etc.) and we are evaluating the idea of initiating additional user fees. I realize that this thread has been carried for quite a while in this group, so thanks for the patience. Because this could be rather a contentious proposal for the department users, I'd like to hear from as many of you who have or had such user fee programs in place. Thanks very much!
Dwight U. Beebe Institut de recherche en biologie vegetale beebed-at-ere.umontreal.ca Universite de Montreal Voice:514-872-4563 4101, rue Sherbrooke est FAX:514-872-9406 Montreal, PQ H1X 2B2 Canada
In regard to general info on user fees - I think that there was a survey a few years ago through MSA (maybe the Education Committee or Tech Forum?)
depending on the size of your operation, and whether you are the only EM facility on campus, you can charge for everything (which can make people feel like you ar nickel&diming them to death) or roll the cost of coaters into the SEM beam time, etc; a two tiered scope charge for assisted vs unassisted usage is an incentive for folks to learn how to operate the beast - but be sure you retain the right to decide who is qualified for unassisted usage; a third level, where you are performing the work might be the same as the assisted usage charge or different, depending on your philosophy
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An important point to ponder in regard to the question of different rates for different users - be very careful ....... most of us are on federal funds and many of the instruments were purchased on federal funds - therefore there are some "rules" that have to be followed - these are the ones I am aware of - there are probably others (if anyone knows where these are concisely enumerated I would like that info)
- If the equip was purchased through government funding 0r is supported by govt funds and you are offering services to the "general public" (industry, hospitals etc), you cannot undercut the prices of any locally available commercial service- check the yellow pages for labs in your area (clinical & industria testing - few of these folks are in MSA)
- Anyone who's research is funded through the feds (don't know if this also applies to state grants) has to be charged the same price - there are ways to give the folks in your dept a break if the dept is providing financial support to the facility, but be sure it is clearly spelled out
- An additional charge = the % overhead your institution takes from fed research grants may be added to the fee for academics from other institutions who are grant funded
- there are also restrictions on using depreciation as a charge basis if the instrument was bought on govt funds and the users are govt funded
- i am sure there are more rules than this but these are the ones I have stumbled across
maybe one of the msa committees should undertake an investigation into this question as we all seem to stumbling around in the dark - to my knowledge no one has yet gotten in hot water on an audit, but as more and more of us are going (at least in part) to a charge basis, it might be a good idea to have some guidelines
1. Our laboratory servers approximately 80 major users (repeat more than 2X a year). 9 from our department, 41 from other departments within our institution, 24 from outside our institution (4 from industry), and 6 from foreign countries.
2. We charge less for departmental users because our department pays part of the service contract on the instruments.
3. We do not charge less based on user expertise. We provide initial training in any aspect of microscopy the user desires. After that we charge a fee for technical time if one of our staff is involved in the microscopy.
4. As outlined in a previous communique (Nestor- do we have a FAQ section?), we breakdown our fees for each individual technique done and cKcharge what it costs us to complete the task.Determining these fees is a major headache, but simplifies administration later and avoids hurt feelings since all are charged based only on what they use. We of course do not nit pick (i.e. charge for each grid, each cc of osmium, etc). Rather we have an average cost for negative stain, cost for cryosectioning, cost for cyroEM, etc.
5. Microscope usage is on a sign up reservation basis. Inexperienced users can only sign up during hours when resource staff are available for consultation. Once we (as a group) are convinced a user can handle themselves we make the microscopes available to them 24 hours a day. Rarely, but it has happened, we encounter someone who is either incapable or unwilling to use the laboratory in a responsible way. These people are denied all access.
6. Marcelle Gillot's summary of the problems of determining users fees is excellent and is almost identical to the constraints we use for determining fees. However, our legal department has a slightly different interpretation of allowable charges for outside, non-grant supported work. They advise that it is allowable to undercut the cost of locally available commercial services as long as you can document (and that may be a problem) that you do so while still recouping all costs of the work without relying on subsidies (goverment grants, institutional support for service contracts, salaries, building maintenance and rent etc.) to cut your cost. In other words, if you are willing to take less profit margin or pay your labor sweat shop wages you can charge less than your commercial competition. You just need to prove that you and your competition are playing on a level playing field.
I hope this is helpful. I would support a movement within MSA to survey and document the issues involved with fee for service operation. Business is clearly something most of us where not traine in and really don't want to spend our time on- however the world being what it is today.......... Regards Jay Jerome
Jim Romanow asks about techniques to produce "black backgrounds"
To possibly state the obvious, you could digitize the images (say using a standard flat bed image scanner ~$1500) apply a digital mask to the area you want to preserve and then "Paint" the background black. That technique is used by commerical artists all the time, many of which are going digital. I know of some "Electron Microscopy Artist's" who basically do the same thing using for example Adobe Photoshop on a MacIntosh.
If you want to go analog, then let me assume that you have tried all the usual tricks like black level shifting gamma processing and other various electronic signal manipulations and they don't get you the image contrast you want.
Have you tried exploiting the directional nature of Backscattered Electron Imaging? Here's what I'd try if I were really pressed to make the background signal as low as possible.
1.) Use a Low Z stub/base this minimizes the BSE signal 2.) Mount the sample on a fine wire stand. 3.) Tilt the stub/holder so that it is tilted away from the BSE detector 4.) Bend the support wire so that the sample faces the BSE detector and so that the sample is between the wire stand and the detector. 5.) Turn off the secondary electron collection field and put a negative bias on the collection grid.
Using this configuration, most secondary electrons from the "background" stub will be rejected by the BSE detector field (I'm of course thinking of a standard PMT detector with a negative retarding field used to reject low energy secondaries) and the BSE from the stub/holder will be for the most part going in the "wrong" direction hence will contribute neglegibly to the signal. You should get a very dark background signal here, however, all edges etc will be enhanced on your specimen and your image may not show the features that you are more comfortable seeing using conventional secondary imaging.
You could of course get fancy with more digital technology and some image processing alogrithms. For example, if you want to bring back the average signal level, record a normal secondary electron image and :-) digitally add/merge the BSE and SEI signals together on your computer, to restore some of the lost SEI information.
If the Coates and Welter does produce true NTSC TV rate signal you could take the signal directly into a TV rate frame grabber board on a computer and the image process to your hearts content. The problem will likely be that the "TV-rate" image that your SEM generates is probably not NTSC but just a fast rate which appears TV like. Slow scan digital imaging is of course do able, however, it's expensive. but you said that you didn't have a slow scan mode, so that option is not viable for you.
I'd probably go the image masking route using Photoshop. If you have patience then you can get excellent results, but so do those guys with the scissors and paste!
Lets talk EDS Detectors! Within a few months time, we will be adding an EDS detector with a thin window for light element detection capability to an existing TEM (Philips CM12, LaB6 electron source, no scanning coils (STEM), + or - 60 degree tilt stage, double tilt analytical grid holder). We are looking at a variety of detector characteristics and are considering: 1. resolution, 2. crystal area, 3. liquid nitrogen cooled vs. peltier cooled (dry), 4. silicon vs. germanium detector crystals, and 5. thin window characteristics. These aspects of detectors are somewhat linked together so its a matter of determining needs and priorities and convolving them with the available budget. We have 16 years of experience running EDS (7.0 micron beryllium window, also soon to be replaced with a new thin window detector) on an SEM, but the addition of EDS to a TEM, with light element detection, will be new territory for us. We are based in a college of agriculture and study a wide variety of biological samples, soils and minerals, and occasional metallic inclusions. We have been discussing these issues with vendors of EDS equipment to enable us to become familiar with the current product line. Now we want to put some of our reasoning and musings in the above areas to the network for feedback from our colleagues. We hope that some experienced hands in the EDS buisness will take a few minutes to respond with the kinds of helpful comments that only someone who has done real, practical work can give. We apologize for the length of this communication, but with the advent of Peltier cooled detectors and the recent introduction of germanium crystals, there is plenty more to consider when buying a new EDS system these days.
1. Resolution. After years of looking at flabby peaks on our 16 year old EDS system (resolution about 162 eV -at- Mn Ka), naturally our inclination is to go for all the resolution we can afford. This seems to imply using a LN cooled premium quality crystal with a 10mm area. Considering the sensitivity to low energy x-rays (0.1 to 1.0 KeV) possible with thin windows, it seems one would particularly want very good resolution to help sort out K,L, M shell overlaps in this low energy region. For light element (or low x-ray energy) detection, a seperate resolution at Fluorine (e.g., 68 eV -at-F) is usually given and we've seen values from 65 to 115 eV -at-F in currently available thin window detectors. Some say "not to worry" about resolution so much because todays peak deconvolution software will sort it out for you, but is it really that good in this energy range?
2. Crystal area. We are aware that the prevailing wisdom suggests that a 30mm (mm squared) crystal should be used on a TEM (with some loss of resolution?) for the larger solid angle of x-ray collection that they have; necessary for thin samples which generally yield lower count rates that a bulk sample in an SEM would give. We wonder if the LaB6 electron source in our TEM will provide enough beam current to get the count rate up (without frying the sample!) to a reasonable level, meaning maximum accumulation times of usable spectra in, say, 200 seconds? Also, do 10mm crystals come mounted in a smaller diameter "snout" which can be moved closer to the sample to increase collection solid angle?
3. LN cooled vs. Peltier cooled ("dry"). (There was a question on the net in late April about "water cooled" detectors but only one response was offered which explained that these detectors are Peltier cooled and water is simply used to carry away heat from the Peltier device). Tho generally a bit more expensive to aquire initially, the main argument for purchasing a Peltier cooled detector given by vendors (Noran, Kevex/Fisons) is the economy of operating them; no hassle with liquid nitrogen to fill twice a week, so saves time and money. Well, we're running a 10 liter dewar on our older EDS right now, and we use LN for cold traps and freezing procedures almost daily. So another 15-20 liters per week(estimate includes loss due to cooling down transfer dewar) isn't going to break the budget. In fact, I would think an EDS dewar is a better place to store LN than the industrial-grade 160 liter dewar that it comes in which vents off more gas than a pack of ardvarks on steroids. Vendor economy calculations apparently assume that you are using LN exclusively for your EDS dewar so that LN cost, delivery fees, tank rental, and LN technician salary, etc, are not spread out over other equipment that consumes LN, so its a maximum LN expense estimate. The down side seems to be higher initial cost and less resolution tho some figures we've seen for some 10mm Peltier cooled crystals, however, range between 138-149 eV (silicon crystals, Noran, Kevex). Now 138 is not bad, but such a detector lists for about $25,000, the 149 eV for $18,300. And if you go to a 30mm Peltier cooled crystal (Noran, generally recommended for TEM applications, see 2. above) the resolution drops to 155 eV -at- MN (112eV -at- F). Is anyone out there operating a Peltier cooled detector on TEM or SEM? They havn't been around as long as LN cooled ones, so we wonder about performance and reliability over time. They also require a seperate water cooling and circulating system to remove the heat being pumped out by the Peltier device. Any problems or commens about that? How's the resolution , etc.?
4. Silicon vs. germanium crystal. The new kid on the block is germanium. And to look at its resolution and countrate through-put values, one wonders if silicon's days are numbered. For SEM only, Link-Oxford offers a thin window 10mm germanium detector with a resolution of 115 eV -at-Mn at 1,000 counts per second (cps), 65 eV -at-F; it rises to only 133 eV -at-Mn at 10,000 cps, ($22,200). It has another slightly lower cost 10mm Ge crystal wth 120 eV -at-Mn, 68-70 eV -at-F ($20,800). For TEM, Noran offers a 30mm germanium crystal with 129 eV -at-Mn. These resolution values look pretty terrific to us!! Less important to us, but another advantage of Ge crystals, is that they absorb higher energy x-rays much better than Si crystals. Silicon absorption efficiency drops rapidly above about 20 KeV (the unabsorbed x-rays pass through the crystal without interacting); Germanium absorption efficiency doesn't begin to drop significantly until x-ray energies get above about 60 KeV. So good detection of high energy K-shell x-rays is available with Ge crystals. But how about at the very low x-ray energies of the light elements; does germanium do OK there? Does it have a dead layer like Silicon crystals do? Are there reasons why germanium detectors with thin windows are not the choice to make for serious light element analysis? Other than being a bit pricey, what are the drawbacks, if any, to Ge crystal detectors?. They havn't been on the market very long, so perhaps there are the usual problems of maintaining performance and reliability until the bugs are worked out. Other than that, it seems to have that "wave of the future" look to us.
5. Thin windows. We read Mark Lund's helpful, but brief, comments on thin windows in late April and look forward to his upcoming book chapter on the subject ( For the MAS, edited by D. Williams and D. Newbury). There are thins and ultra-thins, depending on how sensitive you need to be to the lightest elements, Boron and Beryllium. We only need to see down to Carbon here so a thin window is probably best for our needs. We are concerned about thin window failure rates, especially on the SEM. Do they need to be cleaned often to maintain good low energy x-ray transmission? What other concerns should we have about their care and operation? As Lund pointed out in his comments, if you are specializing in light element analysis, you really have to look at the over-all EDS system in addition to the window material. We do not intend to specialize in light element analysis, but we do want reasonably good detectability, qualitative and "semi-quantitative" of C, N, and O. But we also need to look a lot at the range Z=11 to Z=30, and occassionally above. Does one need specialized software for light element analysis and does it compromise analysis of heavier elements that may be in the same spectrum?
Enough already! Lets give EDS detectors a bit of network time. I'll compile responses into a single file and pump it back out in a few weeks. Thanks in advance to all who respond to this query.
--
Gib Ahlstrand 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
Any input as to good grey-scale printers, as in TRUE grey scale. I'm interested in one that can be accessed from the Mac environment, not thermal paper, and makes nice transparencies. $$ is of minor concern.
I apologize if this repeats a thread.
Tim Foecke, Metallurgy Division, NIST tfoecke-at-nist.gov
___________________________________________________________________ Randy Nessler rnessler-at-uiowa.edu The University of Iowa Central Electron Microscopy Research Facility
On Tue, 7 Jun 1994, Gib Ahlstrand wrote:
} Lets talk EDS Detectors! } Within a few months time, we will be adding an EDS detector with a thin } window for light element detection capability to an existing TEM (Philips CM12, } LaB6 electron source, no scanning coils (STEM), + or - 60 degree tilt stage, } double tilt analytical grid holder). We are looking at a variety of detector } characteristics and are considering:
My question is, without STEM, how are you planning to position your beam on the area of the specimen that you would like elemental information from? I am embarrassed to ask this question, as I obviously haven't researched this in publications. When I asked about collecting x-ray data while performing diffraction, I was told by a service engineer that it would flood the detector with BSE's. Also, there is the problem of diffraction flooding a rather large area of the specimen, and the selected area diffraction aperture allowing the pattern of information to be viewed, while blocking out extraenous information. Are you just interested in average area analysis? I hope I am not the only one curious about this. This is a little off of the topic, and I hope not to detract from the conversation. We are looking to upgrade a detector on a SEM, so I am very interested in following this discussion. Randy
The ANL EMCenter uses a Tek Phaser IISDX dye sublimination printer for all high quality gray scale (& color) work from computers. It is on an Ethernet backbone and directly usable from your Mac "chooser". The quality and price performance for both glossy prints & transparencies is excellent. Cost ~ $10K for the unit then the price/print for 8x10" $2 B&W, $3 Color add an extra $1 to each if you want a transparency.
I'll bring examples to the computer workshop at this year's meeting in New Orleans if anyone want's to have a closer look at real EM data versus manufacturer's demo prints.
Jim Romanow asked about how to get black backgrounds in SEM images:
I've accomplished that using a few simple specimen preparation techniques. The basic trick is to produce a very smooth featureless background, low Z composition prefered:
1. I've done the following for pollen grains or rust spores (dried down onto glass coverslip support surface from water or ethanol, they just stick naturally), should work with insects too, provided that any paint you use to mount the insect to the glass coverslip does not spread out beyond the edges of the insect so you don't see it in the background. Your pin mount technique should work here too: Mount a cleaned (EtOH rinse, air dried) 1 cm diameter round glass coverslip onto a standard aluminum stub using double stick tape and edge the coverslip with carbon paint to provide a conductive path to the aluminum and for extra adhesion. Mount your insect to the coverslip using a tiny dot of carbon paint, or using your pin mount method. Heavy metal coat on a rotary stage as usual and view in the SEM. In spite of the fact that the glass(silicon & oxygen) coverslip is dense and coated with gold-palladium, the pollen grains are brighter than the background and when printing the neg its easy to drop the featureless background into the black, perhaps with a bit of doging of the subject, if it has a simple profile, to burn in the background. The key here is that the background is featureless so that there are no edges of paint globs or other debris which will light up in secondary electron imaging.
2. Similar to the above, is to use a smooth surfaced double stick tape to mount your samples to avoid using paint (for very small samples).Or as a smooth background for your pin mount technique. Some double stick tape surfaces develop tiny cracks or hole patterns in them as a result of being in the vacuum or whatever and may not provide a feature-free background. Spreading carbon paint with a brush to get a low Z background may work if the paint is quite thin but sometimes it leaves too many "brushstrokes" visible and that detail may show up in the background.
3. Another method to get a fairly good low-Z background with a high-Z coating on your sample: Mount samples (directly or pin-mounted) on double stick tape or with a spot of carbon paint onto a stub that has been previously painted with carbon paint (and dried) to hide the aluminum, so you have a low-Z background. Set up your vacuum evaporator rotary stage to zero degrees tilt if you can: We have an old Ladd rotary tilt stage whose "whirling disk" can be re-mounted to zero degrees tilt, that is, horizontal. Set up the evaporator for an overhead carbon rod evaporation on one set of electrodes; set up another set of electrodes for a low angle metal (Au-Pd) coating of about 7 to 10 degrees elevation. After pump-down, set the stage rotating and do the two coatings, in either order. Very little metal coating will hit the stub surface because of the low angle of coating. A lot of metal will hit the target, tho it may be a bit thin on top of the target, hence the overhead carbon coating to get some extra conductivity on top surfaces. This method has worked very well for imaging seeds from 0.05 to 0.4 mm diameter in backscattered imaging (I wanted even "overhead" illumination that BSE gives) with black backgrounds. Should also work OK for secondary electron imaging (gives typical "side" illumination).
Good luck!!
--
Gib Ahlstrand 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
Stuart McKernan suggested that a set of images be organized for testing of printers. The results for which could be put on display at the Computer workshop at the MSA New Orleans meeting. I'm willing to organize this if there is enough interest. If you want to participate Email me seperately (Zaluzec-at-anlemc.msd.anl.gov)
I will collect a set of images from the MSA/EMC library and then setup a FTPable account where interested parties can download the files.
My first impression is that we should have the following in the collection for testing.
SEM-gray scale - Life Science (a bug &/or plant) SEM-gray scale - Physical Science (a computer chip &/or fracture surface) SEM-gray scale - BSE Channeling Pattern
TEM-gray scale - Life Science (cells & collected) TEM-gray scale - Physical Science (Dislocations & Grain Boundary) TEM-gray scale - HREM image of a crystal TEM-gray scale - Electron Diffraction SAED TEM-gray scale - Electron Diffraction CBED
I'm open to more suggestions for test images, make them off line and I'll collect the responses over the next 2 weeks or so. I'll then post the results to the mailing list for further comment before we actually do anything else. I'll attempt to pick micrographs which push the limit of the printers (max contrast & resolution) rather than just a pretty picture. I'll also make sure text is present on each micrograph so that users can also judge the capabilities of the printer to reproduce high quality text as well as an image.
} I think it would be worthwhile to set up an informal meeting related to } this list server. Of course there will be folks coming to the computer } lab, but do you think there would be a place for a presentation of the } server and handouts of how to subscribe/unsubscribe, access, etc? This } might be a chance for exchange of this kind of information, or a way to get } some of the interested parties together. Just a thought.
There will be a tutorial session at the MSA meeting entitled
" A HitchHiker's Guide to Microscopy & Microanalysis Using Telecommunications,Email and the Internet"
by "some guy who wears a hat"
At it I'll have a bunch of handouts about the listserver, the EMMPDL, MSA BBS, a run down of other aspects of Internet (EMail, Telnet, newsgroups, FTP, Mosaic, ......) and anything else I can fit in given time & space & my fortitute. We also plan on-line demos of each if the phone lines can handle the links.
In addition, John Mansfield will be also doing a Tutorial on Image Processing using NIH Image as an example.
These tutorials are set for Thursday Afternoon August 4th, from 1--} Closing. (i.e. ~ 3-4 hours), depending on the audience.
Jay Jerome & John Posthill get the credit for twisting John's and my arms into doing these tutorials. I guess they figure there might be as many as a dozen people that might be interested, anyway, I know of at least 2 people that will be there. So Listserver subscribers, you're welcome to stop by, it's as good a time as any.
The workshop has a reasonably large room again this year, and will be set up for informal as well as formal presentations throughout the week. Of course, I'm more than willing to participate in an even more informal get together at any local pub, so John C. just set a time & place! ;-)
} } The workshop has a reasonably large room again this year, and will be set up } for informal as well as formal presentations throughout the week. Of course, I'm } more than willing to participate in an even more informal get together } at any local pub, so John C. just set a time & place! ;-) } } Nestor Zaluzec } ANL EMCenter } FAQ- Who's buying? ;-} Jay Jerome
As various contributors have pointed out, a black background depends on having the highest possible difference in Z between foreground and a {featureless} background.
With biological samples it is useful to expose the specimen to Osmium (fix or postfix in osmium tetroxide for wet specimens, expose dry specimens to osmium tetroxide vapour in a sealed container for a few hours) and then to mount on a stub coated with gold size.
Japan Gold Size (the stuff that artists use, obtainable from any art supply shop, we use Winsor & Newton brand but others would also be ok) is a sticky liquid, containing mostly linoleic acid polymers. A very thin layer of this on a stub becomes very tacky within 5 or 10 mins at 50 deg C, depending on the characteristics of the batch, and can then be used in the same way as double-sided tape. The thin layer on the stub is made by wiping a slight smear of the liquid over the face of the stub. This soon polymerizes into a glassy, featureless, nonsticky, non-outgassing, low Z layer.
The polymerization can be hastened by a further period at 50 deg C. If your specimens are heat sensitive this further bakeout is not necessary, the process will go to completion at room temp within a few hours. Sputtercoat the sample with a minimum of gold and the high Z osmium in the sample on the smooth low Z gold size will give a nice dark background. Even a very thin layer of gold size on the stub is thick enough to keep the beam away from the relatively high Z stub.
The gold size idea is not our own, it was published long ago in a paper that we cannot now find, but it works really well for a wide variety of specimens. In addition it is cheap, natural, biodegradeable, nontoxic and apparently also non-allergenic! Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
Free to a good home. Philips EM 300 Electron Microscope now surplus to requirements due to rationalization of Faculty EM Services. Very Good Condition. Philips maintain the instrument. Further details: Mr J N Brown, Department of Physiology, University of Edinburgh, Teviot Place, Edinburgh, UK. Telephone (031) 650 3273, fax (031) 650 6527.
Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable To: Microscopy-at-anlemc.msd.anl.gov
=46ollowing the question from R. Nessler: My question is, without STEM, how are you planning to position } your beam on the area of the specimen that you would like elemental } information from? I am embarrassed to ask this question, as I obviously } haven't researched this in publications. When I asked about collecting } x-ray data while performing diffraction, I was told by a service engineer } that it would flood the detector with BSE's. Also, there is the problem } of diffraction flooding a rather large area of the specimen, and the } selected area diffraction aperture allowing the pattern of information to } be viewed, while blocking out extraenous information. } Are you just interested in average area analysis? I hope I am not } the only one curious about this. This is a little off of the topic, and I } hope not to detract from the conversation. We are looking to upgrade a } detector on a SEM, so I am very interested in following this discussion. } Randy
In the CM12, you can work under normal TEM observation and reduce step by step the size of the illumination spot down to about 20 nm while still retaining enough beam current in the probe to perform EDS microanalysis in a reasonable time. For that purpose you will use the spot size knob (physically the 1st condenser lens). This way is available on most modern TEMs too. DON'T FORGET TO REMOVE THE OBJECTIVE APERTURE, which otherwise will produce a hudge amount of X-rays and dazzle the EDS detector (it may take some minutes to fully recover its normal dead time!). It can be helpful to switch to diffraction observation if you want to analyze a tiny precipitate that give a different diffraction pattern than the surroundings. Be careful, a probe shift may happen due to the weak coupling of magnetic fields of the diffraction lens and the objective lens. So you will have to slowly shift (blind) the probe until you find again the caracteristic diffraction pattern of your phase. Then start microanalysis. If you see the pattern changing during the analysis, you should stop acqisition, shift the probe again and then continue the analysis. I desagree entirely with your service engineer. Switching to diffraction do not change anything to the magnetic field around the sample (ie, the objective lens) within the limit of the weak coupling mentioned above. So there are no more BSEs reaching the detector in one mode than in the other. Thus there is no reason not to use diffraction instead of image if you feel easier to decide wether or not you are firing on the right phase. He was probably thinking to the observation in low magnification mode (LM mode for Philips) where the excitation of objective lens is switched off or very strongly reduced. Then BSEs do no longer spiral in the magnetic field and then are allowed to escape toward the EDS detector.
I hope this will help you. Do not hesitate to contact me again if I wasn't clear. But I am leaving Friday evening for the next two weeks. Best regards
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Institut Interd=E9partemental de Microscopie Electronique Address: EPFL-I2M, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 (Central European Time) Fax: +41(21)693 44 01 ______________________ Eudora 2.0.2 __________________________________
} ___________________________________________________________________ } Randy Nessler } rnessler-at-uiowa.edu } The University of Iowa Central Electron Microscopy Research Facility } } } On Tue, 7 Jun 1994, Gib Ahlstrand wrote: } } } Lets talk EDS Detectors! } } Within a few months time, we will be adding an EDS detector with a thin } } window for light element detection capability to an existing TEM (Philips } } CM12, } } LaB6 electron source, no scanning coils (STEM), + or - 60 degree tilt stage, } } double tilt analytical grid holder). We are looking at a variety of detector } } characteristics and are considering:
Firstly, thanks for the useful info on EDS detectors. One question: Has anyone had success with annealing detector crystals to combat incomplete charge collection? } } My question is, without STEM, how are you planning to position } your beam on the area of the specimen that you would like elemental } information from? I am embarrassed to ask this question, as I obviously } haven't researched this in publications. When I asked about collecting } x-ray data while performing diffraction, I was told by a service engineer } that it would flood the detector with BSE's.
Switching to diffraction mode focuses the intermediate/diffraction/P1 (nomenclature depends on manufacturer) lens onto the back focal plane of the objective. It does (should) not influence the illumination seen actually at the specimen. Maybe your service engineer is thinking of fried (jellied?) EELS detectors ? :-) (which company is he from, btw?). I've lost the post now (hereafter referred to as the "lost-poster"), but someone mentioned weak coupling between INT and OBJ lenses - surely this problem, re spot shifting, is not a problem with correct rotation centring. Life starts getting very tricky, however, when your specimen is charging/magnetic/unstable - I won't go into details here though, since that's another issue. For precipitates/inclusions/nano-crystalline materials, viewing the diffraction pattern is indeed a good way to check the spot position, as already pointed out. Quantitative microanalysis of these features is a headache, though, but that's also another thread.
} Also, there is the problem } of diffraction flooding a rather large area of the specimen,
Use a smaller spotsize (C1 and C2 controls). CBED often is performed with tiny (~1nm or less FWHM) spot-sizes. LaB6 or FEG will help here. In a darkened room, you should be able to see sufficient information in the image formed with the smallest C1 and slightly defocussed C2 to orient the specimen. Chances are that if you cant see anything, your count rate will be too low anyway.
and the } selected area diffraction aperture allowing the pattern of information to } be viewed, while blocking out extraenous information.
Not sure what you are getting at here. The SAA, being far down the column (position varies - any post-specimen image plane will do), is well removed from anything to do with EDS, although possibly it may contribute X-ray counts if sufficiently high. It's a simple matter to find out if it is contributing and whether you need to make sure all users remove it prior to analysis. As the lost-poster said, it is *very* important to remove the Obj Aperture (OA) or you may start encountering incomplete charge collection (which manifests itself as an asymmetric, low-energy tail to each peak) soon afterwards. :-(
} Are you just interested in average area analysis? I hope I am not } the only one curious about this. This is a little off of the topic, and I } hope not to detract from the conversation. We are looking to upgrade a } detector on a SEM, so I am very interested in following this discussion. } Randy
The comments above refer to TEM/STEM. Life is quite different for SEM. We too are considering SEM detector upgrade/purchases so are also v interested in this thread (re detector crystals).
I'm working on analyzing dislocation arrays in ferroelectric PbTiO3 thin films. I'm searching for any software, preferably for Macintosh or DEC Alpha UNIX, that will simulate dislocation contrast under various diffraction conditions. I'm aware of ONEDISANL and TWODISANL (available through the EMMPDL archive), but would like to try other options if they are available. Suggestions?
Thanks in advance.
Stephen Streiffer Max-Planck-Institut f. Metallforschung Stuttgart, Germany
In the TEM literature there is a technique called pseudo-replica formation where formvar or collodion is poured in a thin layer over a surface and alloed to dry. It is then floated off, havin trapped particulates and large molecules which can then be observed with TEM. Has anyone done or does any one know of a similar technique for SEM. I need to enumerate particles attached to moist surfaces that cannot be processed by conventional means for SEM (living skin). I wonder if pressing scoth tape to the surface would pick up all the particles down to the several micron range???
We're seriously considering starting a user-fee type of arrangement for our electron microscopy facility. I was wondering if and how much other similar facilities charge for use of microscopes and equipment as well as consulting, etc..... Our facility is staffed by a faculty advisor and a half-time facilities manager.
I would even be willing to run a survey and compile the results if that is what it takes to get this information.
Thanks,
Todd Voiles Central Facility for Electron Microscopy Center for Materials Research and Analysis University of Nebraska, Lincoln, City Campus
} Hi, } } } We're seriously considering starting a user-fee type of } arrangement for our electron microscopy facility. I was wondering if } and how much other similar facilities charge for use of microscopes } and equipment as well as consulting, etc..... Our facility is staffed } by a faculty advisor and a half-time facilities manager. } } I would even be willing to run a survey and compile the results } if that is what it takes to get this information. } } Thanks, } } Todd Voiles } Central Facility for Electron Microscopy } Center for Materials Research and Analysis } University of Nebraska, Lincoln, City Campus } } tvoiles-at-unlinfo.unl.edu } Todd: Here at UMBC we charge for EM services but since we are a state institution we can't make a profit. All of our charges cover just the costs. We are probably on the low end as far as charging for services go. If it will help, send your fax number and I will fax a copy of our charges to you. Phil Rutledge Phone: (410) 455-3582 FAX: (410) 455-3875
A minor point, but potentially an important one for the future of Microscopy Listserver.
The Microscopy Listserver, and the MSA BB system are not the same entity. Although I basically run both (and also the EMMPDL, & Coordinate the MSA Computer workshop), the funds for each do not come out of the same pot.
To most of you things are transparent and (relatively?) seemless. I've tried to keep it that way intentionally as I've tried to make sure that everyone has access with the maximum amount of versatility (of course not everything works like it's supposed to but we're getting there slowly). The reason for this note is that I've seen some references lately where these admittedly subtle distinctions are accidently blurring, for example, in references to where things are located or how to get access to some information. This may not seem like a big deal to most of you, however, ultimately, I do need to document things to present and future funding sources (i.e. there is no free lunch) so when and if you talk about these different systems please try acknowledge them as different beasts.
Just for the record:
The MSA Computer Workshop is funded totally by MSA, except during joint society meetings, when our sister societies chip in a proportion of the costs. It is a week long interactive workshop held each year at the annual meeting of MSA.
The MSA BBS is jointly funded by MSA and ANL EMC. It is a regular BBS system runing on a PC with both Internet and Telephone links. MSA supports an 800 number (1-800-MAS-EMSA) as well as having purchased the PC and some of the software. The ANLEMC pays local costs (power, phone lines, network lines) and spends time & effort to keep it updated and running.
The EMMPDL is mainly funded by the ANLEMC, MSA contributes toward some expenses when local/sister societies want copies, or when the library is distributed at meetings (for example in the MSA computer workshop at New Orleans, or local society meetings).
The MICROSCOPY LISTSERVER (i.e. this EMail distributed discussion group) is run by by the ANLEMC but is not "officially" supported yet by any funding agency, basically I started it on the side as an experiment (call it a voluntary service to the microscopy community). Right now the ANL EMCenter ultimately pays the network costs through it's general overhead costs (so ANL and DoE should get some credit).If it is referenced, especially if you want to acknowledge it please simply use the phrase "The ANLEMC Microscopy Listserver". Try not to confuse it with the MSA BBS, while I'll know what you mean, if I need to raise support funds in the future (and I will need to) then it will need to be acknowledged appropriately.
Those of you who are reading the Microscopy Listserver via Newsgroups are getting a mail feed from ANLEMC, via a mailserver at the University of Michigan arranged by John Mansfield. Also,if you are exclusively on a newsgroup system you should know that the newsgroup replies do not presently filter back to the Microscopy Listserver subscription list. This means that your replies and/or questions do not reach most of the users of this system. This is on the list of things to fix, but it's one of those things that takes time and $$... If you want to reach the maximum coverage, you should post to:
We've just received an Alden 9315ctp continuous tone printer, but we were faced with a surprinsigly poor information for operating the printer, specially having in mind its price. The only executables they send are a tprnt.exe, presumably for TIFF files (no indication for the version), and a couple of conversion executables like t2r.exe and x9pc.exe. Unfortunaly in our hands these programs seems to have quite an random behaviour, either printing the image correctly, either producing truncated, layed, or tottaly corrupted images. So far we've tested a number of programs to generate the TIf's and Raster's (PSP, XView, PStyler, etc.) and so far we could not find a pattern for this behaviour. Being in a small european country, the local rep's can't be of much help (actually it's the first one they sold). Therefore I would to ask help to anyone who could provide me with any info on the operation of such printer, including other drivers for direct printing from the software (Windows for instance), info on net sharing drove by UNIX machines, and any possible internet contact with the guys from Alden. Thank you very much for any help.
________________________________________________________________ Jose A. Feijo Dept. Biologia Vegetal, Fac. Ciencias Lisboa Ed. C2, Campo Grande, P-1700 LISBOA, PORTUGAL
I am working on CaM localization and quantification in plant cells, and for that I have been using polyclonal antibodies raised in rabbit against bovine CaM. These antibodies were sold by SIGMA, but now they only sell MAb's, which does not detect the plant CaM.
So is there anybody out there with a stock of polyclonal anti CaM. It does not matter if the antibodies are raised against plant or animal CaM, but i would prefere if they were raised in rabbit.
Gred Erdos was inquiring about making replicas for SEM of living skin
One method you could try is to use dental molding compound - (the stuff they use to make impressions for orthodontics etc) - you mix it up, press it onto the surface of interest and it hardens up very quickly and can be removed without damaging the original surface -
We have done this a a class exercise - using the replica as a mold to make a "positive" with Spurrs resin - we generally did 2 applications, the first being used to remove the small surface debris which you want to look at so I suspect it should work for you
The stuff we used was called "Cuttersil" and was available through Columbus Dental PO Box 620 St Louis Mo 63188
} } *Received through multiple hands across the sea and along the internet } } } } INTERNET VIRUS ALERT! Date: Wednesday, May 25, 1994 } } } } A Virus has been discovered on Internet that is disguised as CD-ROM shareware. } } } } Unknown hackers have illegally put the Chinon name on a destructive shareware } } file and released it on the Internet. This catastrophic virus is named } } "CD-IT." --DO NOT DOWNLOAD. IT WILL CORRUPT YOUR HARD DRIVE. The program, } } allegedly a shareware PC utility that will convert an ordinary CD-ROM } } drive into a CD-Recordable (CD-R) device, which is technically impossible, } } instead destroys critical system files on a user's hard drive. The program } } also immediately crashes the CPU, forces the user to reboot, and stays in } } memory. } } } } Widest dissemination is requested. } } } } } } } } (Mr) Lindsey Thomas Martin (lmartin-at-sfu.ca) } Dialogue: Canadian Philosophical Review } Vancouver Studies in Cognitive Science } International History Review } Simon Fraser University, Burnaby BC Canada. }
---------------------------------------------------------- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego CA 92182-0057 phone: (619) 594-4523 fax: (619) 594-5676 email to sbarlow-at-sunstroke.sdsu.edu
A visiting scholar at the Friday Harbor Labs is looking for parts for a JEM transmission microscope (model #100B). If anyone knows of a source please reply to Scott Schwinge at schwinge-at-fhl.washington.edu
Just so you know, part of the problems which caused a large number of Undeliverable mail messages has been cured. A server at the Univ. of Michigan had gone batty and was fixed by John Mansfield. We still have another server somewhere in the system which cannot deliver some mail to "previously" valid addresses. I'm investigating the problem.
___________________________________________________________________ Randy Nessler rnessler-at-uiowa.edu The University of Iowa Central Electron Microscopy Research Facility
On Wed, 15 Jun 1994, MICHAEL DELANNOY wrote:
} I am looking for software that will drive a confocal microscope but doesn't } save image files in the .pic form but something more useful for other } Mac imaging programs such as Photoshop. Alternatively (and more likely) a } program that converts .pic to .gif or PICT without loss (at least significant } loss) of image quality. The clumsyness of manipulating images for figures in } comos is not aceptable } } } } Shawn Burgess } c/o Mike Delannoy
Talk to Biorad about the program .pic to .tiff. We have it on our MRC-600. Lately, people have been copying files to floppies and taking them to Adobe Photoshop. They are able to manipulate the .pic images knowing that the header is 76 bytes, there are 768 columns, 512 rows, and bytes/pixel = 1. Granted, it is a little cumbersome to do this. It appears that you have the confocal and Mac already, and is proably most economical to research the means to interface/import images. Randy
...... am looking for software that will drive a confocal microscope but doesn't save image files in the .pic form but something more useful for other Mac imaging programs ....
You should seriously look into getting a copy of NIH-Image. The current version level is 1.55 and 1.56 is in beta. You may download a copy from zippy.nimh.nih.gov using anonymous FTP. Alternatively if you attend the Microscopy Society of America Meeting in New Orleans in August you may get a copy at the Computer workshop & software exchange.
Message-Id: {MAILQUEUE-101.940615150052.256-at-vanlab.paprican.ca} To: Microscopy-at-anlemc.msd.anl.gov
Are there any microscopists out there who've worked on morphology differences between wood species? We're trying to find data on northern and interior British Columbia wood species with respect to fibre dimensions. If anyone has any insight out there I'd be grateful to hear from you, perhaps directly since this is a topic of extremely narrow interest and needn't tie up the listserver (unless someone else wants the info too; let me know). This is also kind of a shot in the dark to draw some of you forestry related microscopists out of the woodwork (pun unintentional). Thanks.
James Drummond Pulp and Paper Research Institute of Canada Vancouver, B.C.
First let me say that I AM NOT LOOKING FOR A JOB! But I would like to see a discussion of the future of technical jobs in EM. I always look at the classified ads in the EMSA (now MSA) bulletin. I have always found it interesting to see where tech jobs were and if my qualifications were keeping me competitive with the market. The last few MAS bulletin didnot have many, if any, EM job.
I have had scientist out side EM tell me that EM had had it peak, as all tecnologies do, and now has less and less uses. I believed that although pure ultrastructual studies of biological tissue has been pretty well covered, new advances would renew the need for trained people.
Is it true that EM is past its prime? EMSA did take out the "E"
Are people advertising job some where else?
Is money so tight that EM has become to expensive to be widely used?
Are service facilities taking the place of numerous smaller labs?
Are old EM tech. so full of Glut and OsO4 that they are not dying off?
Someone please give me some comfort. I still have 28 more years until retirement.
We will be purchasing a pc operated SEM with the intent of being able to remotely control the instrument. Initially, the remote site will be in our building and then a second phase is to be able to remotely control the instrument from a sister campus. Is anyone doing remote control of a SEM? If so, could you address the following questions?
Have you been able to accomplish remote control other than serial?
What functions can you perform?
What software interfaces have you used to access the SEM from elsewhere? (carbon copy, pc anywhere, etc).
What are the speed considerations?
Have you attempted to send both video and the data signals in real time?
How have you accomplished it?
If you have not accomplished this task, do you have any ideas or suggestions about how it could be done?
In times of very tight money, I don't think any of us can provide much comfort. However, I think Larry's points are good ones. I do think more of us are employing light microscopy (or going back to our original roots in LM). I think this is good because often LM/EM combined studies can be much more powerful than single technology. EM provides the resolution, but LM provides greater sample area and dynamic information. Our scheme to survive (contact me again in 5 years to see if it works), is to attract non-traditional EM users (biochemists, physiologists, etc) by helping design their experiments and making use of EM as easy as possible. All that is required of them is the ability to focus and capture the images. This has worked well for us over the years, the biochemistry department rather than the anatomy or pathology department constitutes our biggest users. It takes more of my time but it keeps us solvent. So, yes ithink there is still a large need for us dinosaurs (molecular genetics still won't answer all questions and a picture is still worth a thousand words even with inflammation) but you have to be more of a slalesman and gear your services to the community needs. My humble opinion.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Pathology * * Bowman Gray School of Medicine of Wake Forest University * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
On Wed, 15 Jun 1994, Larry Hawkey wrote:
} } First let me say that I AM NOT LOOKING FOR A JOB! } But I would like to see a discussion of the future of technical } jobs in EM. I always look at the classified ads in the EMSA (now MSA) } bulletin. I have always found it interesting to see where tech } jobs were and if my qualifications were keeping me competitive with the } market. The last few MAS bulletin didnot have many, if any, EM } job. } } I have had scientist out side EM tell me that EM had had it peak, as } all tecnologies do, and now has less and less uses. I believed } that although pure ultrastructual studies of biological tissue has } been pretty well covered, new advances would renew the need for } trained people. } } Is it true that EM is past its prime? EMSA did take out the "E" } } Are people advertising job some where else? } } Is money so tight that EM has become to expensive to be widely used? } } Are service facilities taking the place of numerous smaller labs? } } Are old EM tech. so full of Glut and OsO4 that they are not dying off? } } Someone please give me some comfort. I still have 28 more years until } retirement. } } Larry Hawkey } Hawkey-at-neuro.duke.edu }
EM certainly has a future if our experience is any indication. We run a central service lab and we have never been without work to do for more than half a day. Seems just as we catch up (which rarely happens) the phone starts to ring.
We may not be seeing positions advertised for tech's due to the time lag involved in the publication of professional society newsletters. When I have a vacancy in the lab it usually occurs with little more than a few weeks notice and I need to fill that spot quickly and cannot wait for the next MSA Bulletin to come out. So I always turn to Jungle Drums for faster communication. ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
} } First let me say that I AM NOT LOOKING FOR A JOB! } But I would like to see a discussion of the future of technical } jobs in EM. I always look at the classified ads in the EMSA (now MSA) } bulletin. I have always found it interesting to see where tech } jobs were and if my qualifications were keeping me competitive with the } market. The last few MAS bulletin didnot have many, if any, EM } job. } } I have had scientist out side EM tell me that EM had had it peak, as } all tecnologies do, and now has less and less uses. I believed } that although pure ultrastructual studies of biological tissue has } been pretty well covered, new advances would renew the need for } trained people. } } Is it true that EM is past its prime? EMSA did take out the "E" } } Are people advertising job some where else? } } Is money so tight that EM has become to expensive to be widely used? } } Are service facilities taking the place of numerous smaller labs? } } Are old EM tech. so full of Glut and OsO4 that they are not dying off? } } Someone please give me some comfort. I still have 28 more years until } retirement. } } Larry Hawkey } Hawkey-at-neuro.duke.edu } Larry,
My own bias is that EM use for biology will increase in the future. We have all seen a very striking renaissance of morphology at the LM level over the last few years. For example, everyone in molecular biology now wants to do immunocytochemistry, in situ hybridization and to localize reporter gene expression. And molecular biologists are finding out that they can utilize these methods intelligently only if they have some understanding of microscopes, tissue organization, cells, ultrastructure and basic morphological methods and strategies. New technologies are greatly expanding the capabilities of morphological methods. For example, the confocal LM can show where fluorescent ICC activity is in cells with much greater clarity than previous fluorescent microscopes could. All this is a far cry from a few years ago, when graduate students thought that if they learned how to clone genes their futures would be assured.
I really believe that these trends will continue, and will extend more and more to the EM level. It is often important to localize antigens within cells, which can be done by immunocytochemistry with gold particle labels. When lacZ (yielding beta galactosidase) is used as a reporter gene, it is often very difficult to identify the blue cells seen with the Xgal reaction in a complex tissue. In that case, localizing the Xgal staining reaction at the EM level (where it can be seen as dark blobs in the cells) allows one to utilize ultrastructural critera for the cell identification. There are many other examples.
My own convictions along these lines are clear from the fact that for the last two years I have offered a graduate course, "Morphology for Molecular Biologists" here at the University of Michigan. The course is held once a week (a three hour session), and includes lectures and demonstrations covering LM, TEM, SEM, a 2-hr "minicourse" in histology, ultrastructure, and then immunocytochemistry, autoradiography, in situ hybridization, localization of reporter gene expression, and finally confocal LM. Grad students from various departments and schools at the U-M have seemed highly motivated in the course.
So I am betting that the electron microscope does indeed still have a future.
A. Kent Christensen Department of Anatomy and Cell Biology University of Michigan Medical School akc-at-umich.edu
Microscopists: I operate a core facility here at UMBC and we are continually getting new contracts for EM services. I even get requests from universities that have EM facilities on campus. EM did take a nose dive but I think EM is on the way back up. I know this may cause an argument, but even molecular biologists are starting to see that instead of looking at gels all the time for information, additional and important information can be found by doing EM. Yes, I do have a bias against molecular biologists. When you have been constantly told by molecular biologists that there is no future for EM, you sort of get biased. I guess they forget that molecular cell biology is just another TECHNIQUE as is EM. Have a cold one for EM! Phil
The future of EM also concerns me. Molecular biology is in it's prime and the investigators seem to be following the trail of grant money that follows the latest technological fad. Who can blame them? However, I am still optimistic about the future of all microscopy. Similar to other labs, we have expanded our LM capabilities and entered into confocal microscopy. Further, we are also responsible for flow cytometry. Contracts with industry have been very important to the lab's fiscal health. Diversification enables us to select the most relevant technology for the client's needs. Correlation of molecular work to the morphology will bring the investigators back--in a different way. Immuno, fluorescence, digital imaging/analysis etc., are all going to be a part of it. Look at the environmental SEM, or SFM, or AFM. Actually, there is a lot to be excited about. What do the material microscopist's think? Eh, Nestor?
************************************************* _______________________ * * | | * * | _____ ______ | * Charles J. Butterick (Chuck) * |__| | | |__| * Electron Microscopy Center * | | * Department of Cell Biology and Anatomy * ______| |______ * Texas Tech University Health Sciences Center * | __ __ | * Lubbock, Texas 79430 * |__| | | |__| * USA * | | * Phone 806 743-2706 voice * | | * 806 743-2707 fax * | | * Email hecub-at-ttacs.ttu.edu * | | * * __| |__ * * |_______| *************************************************
We would like to record some information with our images, perhaps in a header. It is getting to cumbersome to keep matched doc files with their images. We would like to be able to move and store one file per image and have that file also contain all of the documentation, instrument parameters, etc. We would like that information to be in ASCII so that it can be read easily.
I'm looking into using a TIFF tag such as imagedescription, to designate a long string or narrative that would describe the image. That string might look something like this: ((operator Dave Bright)(Instrument JEOL 8500)(Magnification 1000x 5=B5 field width)(date 16 June 1994)(Image_mode transmission bright field)(comment Sample #3 lot 445x2. 100nm gold coated after fine polishing. etc. etc.))
I presume we would have to write a small application that would append/edit/extract this part of the TIFF file. Other tiff file readers such as NIH Image would ignore it - the actual text would probably be put after the image, although this should not make any difference.
Anyway, for you image documenters out there: How do you do it? Are there already utilities around for keeping information along with the images, other than database type utilities?
Thanks :8o) Dave
------------------------------------------------------------- David S. Bright bright-at-enh.nist.gov Microanalysis Research Group Bldg. 222 (Chem.) A113 National Institute of Standards & Technology (NIST, formerly NBS) Gaithersburg, MD 20899-0001 / USA 301-975-3911 (voice), 301-216-1134 (fax) "A false balance is an abomination to the Lord but accurate weights are his delight.", Proverbs 11:1
} } This is hardly microscopy but rather a long shot. I have a colleague here } looking for a copy of the TRIM (transport of ions in solids) program, which we } believe to be public domain or otherwise available, for computing } ion distributions inside a solid after ion implantation. Can anyone help } direct us in the right direction? We would prefer a Mac or PC version, } but we also have other workstations so could probably run almost any } version. } } Thanks. } } Tony Garratt-Reed } } }
The author is James F. Ziegler IBM-Research, 28-0 Yorktown, NY 10598
Greetings, A client wants to embed ants in plastic for both light and TEM and has had problems with the exoskeleton embedding adequately in paraffin. We have no experience with insects. Are there procedures for softening the chitin? Any help would be appreciated.
************************************************* _______________________ * * | | * * | _____ ______ | * Charles J. Butterick (Chuck) * |__| | | |__| * Electron Microscopy Center * | | * Department of Cell Biology and Anatomy * ______| |______ * Texas Tech University Health Sciences Center * | __ __ | * Lubbock, Texas 79430 * |__| | | |__| * USA * | | * Phone 806 743-2706 voice * | | * 806 743-2707 fax * | | * Email hecub-at-ttacs.ttu.edu * | | * * __| |__ * * |_______| *************************************************
John Benci asked about suppliers of used microscopes. Since used equipment is my favorite kind I have collected a list of companies that include electron microscopes in thier inventory. Of course this list is not complete, and I would love to hear about any others:
Cusaco Phone 908-502-9246.
Scientific Equipment and instrument Company 408-428-0464
OOPS! Cusaco's phone is 908-502-0246!! ignore the one above!
The Source 1-800-722-7719
Techlink 408-922-0888
International Equipment Trading Company 708-913-0777
} Greetings, } A client wants to embed ants in plastic for both light and TEM and } have no experience with insects. Are there procedures for softening the } chitin? Any help would be appreciated.
You might look at a recently published method in which the cuticular or waxy surface is primed by treatment with gamma-glycidoxypropyl trimethoxysilane prior to resin embedment in LR White. the paper was in Microscopy Research and Technique 21:355-360 (1992). The silane is available from Polysciences,Inc. (800)-523-2575.
} John Benci } Materials Science & Engineering } Wayne State University
} e-mail address: jbenci-at-eng.wayne.edu
} ------------------ RFC822 Header Follows ------------------ } Received: by mse.engin.umich.edu with SMTP;16 Jun 1994 12:29:09 U } Date: Thu, 16 Jun 94 12:15:18 EDT } From: jbenci-at-hub.eng.wayne.edu (John Benci) } Message-Id: {9406161615.AA29719-at-ss0.NIS.ss0} } To: microscopy-at-anlemc.msd.anl.gov } Subject: Used SEM's
A gentleman by the name of Clark Houghton, located in Ohio, would be the best and closest source I know of. Like me, he offers maintenance services on a wide range of electron microscopes. Unlike me, he also deals in used, refurbished instruments and also has leasing options. There are others offering used instruments, but if I were looking for a used instrument, he's probably the only one I would talk to.
Clark Houghton Phone (513) 927-5373 FAX (513) 927-5557
BTW, although I don't generally deal in used equipment, I often help my customers find new homes for their old stuff. None that I know of currently have equipment available, but I expect at least a couple of machines on the market in the next 6 months to a year. If your needs are not immediate, send me email with a more specific listing of your requirements.
Just to clarify - I have no financial relationship to Mr. Houghton, and only a passing friendship. He is well known and respected in the field.
Allen R. Sampson
Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174
Could you please tell me if you are a state or private institution. Who bought the microscopes, who pays the tech salaries and who buys the supplies at priori. Finally who pays for the service contrast of each instrument. Do your charges to customers reflect these categories? How much let us say you would charge for embedding a piece of heart cuttting at three levels and making 30 pictures (5X7 inches) 10 at low mag (1-3) 10 at 5-20 and ten at 20-100? Thanks.
***** ************ ************** ************ *Cesar D. Fermin, Ph.D |Fax (504) 587-7389 *Tulane Medical School |Answ. Mach.(504) 584-2618 *Pathology/SL79 |Secretary (504) 584-2436 *New Orleans, La 70112 | Lab (504) 584 2521 ***** ***************** *********************** _______________________________________________________________________________ Cc: Microscopy Group
In answer to Dwight Beebe's additional questions:
1. Our laboratory servers approximately 80 major users (repeat more than 2X a year). 9 from our department, 41 from other departments within our institution, 24 from outside our institution (4 from industry), and 6 from foreign countries.
2. We charge less for departmental users because our department pays part of the service contract on the instruments.
3. We do not charge less based on user expertise. We provide initial training in any aspect of microscopy the user desires. After that we charge a fee for technical time if one of our staff is involved in the microscopy.
4. As outlined in a previous communique (Nestor- do we have a FAQ section?), we breakdown our fees for each individual technique done and cKcharge what it costs us to complete the task.Determining these fees is a major headache, but simplifies administration later and avoids hurt feelings since all are charged based only on what they use. We of course do not nit pick (i.e. charge for each grid, each cc of osmium, etc). Rather we have an average cost for negative stain, cost for cryosectioning, cost for cyroEM, etc.
5. Microscope usage is on a sign up reservation basis. Inexperienced users can only sign up during hours when resource staff are available for consultation. Once we (as a group) are convinced a user can handle themselves we make the microscopes available to them 24 hours a day. Rarely, but it has happened, we encounter someone who is either incapable or unwilling to use the laboratory in a responsible way. These people are denied all access.
6. Marcelle Gillot's summary of the problems of determining users fees is excellent and is almost identical to the constraints we use for determining fees. However, our legal department has a slightly different interpretation of allowable charges for outside, non-grant supported work. They advise that it is allowable to undercut the cost of locally available commercial services as long as you can document (and that may be a problem) that you do so while still recouping all costs of the work without relying on subsidies (goverment grants, institutional support for service contracts, salaries, building maintenance and rent etc.) to cut your cost. In other words, if you are willing to take less profit margin or pay your labor sweat shop wages you can charge less than your commercial competition. You just need to prove that you and your competition are playing on a level playing field.
I hope this is helpful. I would support a movement within MSA to survey and document the issues involved with fee for service operation. Business is clearly something most of us where not traine in and really don't want to spend our time on- however the world being what it is today.......... Regards Jay Jerome
Does anyone have a procedure for producing skeletal carbon TEM grids with holes sufficiently small to capture particles ranging in size from 200 nm to 30 nm?
Thanks.
-- Diandra L. Leslie-Pelecky Center for Materials Research and Analysis PH: (402) 472-9178 University of Nebraska FAX: (402) 472-2879 Lincoln, NE 68588-0113 diandra-at-unlinfo.unl.edu
I wish to confer with someone knowledgeable about the cryogenic aspects of EDS detectors. e.g., loss of insulating capacity of the Dewar, effects of warmup - quantitative, i.e., resolution with time and temperature- activation energy of the kinetics, effects of operating at dry ice -acetone temperatures.
A really good reference will suffice.
Please reply direct. I will prepare digest of any information that I receive for the list.
Message-Id: {9406201136.AA17097-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Software / Font needed Orig-Author: {mwinton-at-nosc.mil (Michael J. Winton)}:ddn:wpafb ----------------------------------------------------------- I apologize in advance for a post which is somewhat related to microscopy but not very closely. Please skip this message if that bothers you.
I am looking for a font for the Macintosh which has overlined numbers (Miller indices) built in. I have been searching and searching, and am tired of "drawing" the indices and pasting them into my documents. I'm sure that someone out here must have such a font. A public domain font editor would also be useful-- then I could create such a font and make it available to others. I have yet to find a font editor in an FTP site--font utilities are usually useful for little more than cataloging. I thought that there would be enough scientists in this newsgroup who have to deal with this regularly that one of you might know of something.
Please let me know if you can help.
Michael Winton (mwinton-at-nosc.mil / -at-zazen.lbl.gov)
Does anyone have a procedure for producing skeletal carbon TEM grids with holes sufficiently small to capture particles ranging in size from 200 nm to 30 nm?
Thanks.
-- Diandra L. Leslie-Pelecky Center for Materials Research and Analysis PH: (402) 472-9178 University of Nebraska FAX: (402) 472-2879 Lincoln, NE 68588-0113 diandra-at-unlinfo.unl.edu
Dear Light Microscopists, Does anyone know of a specific LM stain for fibrocartilage? A student is doing a study on a part of the skull and joints and would like some help. Any suggestions?
Thank you Piotr Swierczynski
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
Dear EM people, Does anyone know of any company (other Fisons UK) manufacturing gold or gold/palladium ring targets for Biorad sputter-coaters? We have just been informed (after waiting two weeks and having being informed that it was on its way) that we have a 3-4 week wait for a new one from Fisons UK, a slight problem as due to an "administrative oversight" there is no spare. Any suggestions for an alternative source in future would be gratefully received. Yours faithfully RE
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
In Word for Macintoshes, Miller indices are easy to make using the formula editor.
For those using Macs and don't know how to use the formula editor I can send a small Word 5.1 document with some examples of Miller and Weber indices. (BTW Word for Macs is much easier to use than WordPerfect!!).
Cheers, Timon
------------------------------------------------ Timon Fliervoet, Timonf-at-earth.ruu.nl, Faculty of Earth Sciences, Department of Geology, Utrecht University, P.O. Box 80.021 3508 TA Utrecht, the Netherlands. tel: (0)30 - 535054, fax: (0)30 - 537725
SPI Supplies carries the targets you are looking for and they should fit the Bio-Rad instrument. They are located in West Chester, Pennsylvania, USA. Tel # 1-800-242-4774 and FAX # 1-215-436-5755. Good luck!
In message Tue, 21 Jun 1994 17:07:32 +1200, richard.easingwood-at-stonebow.otago.ac.nz (Richard Easingwood) writes:
} Dear EM people, } Does anyone know of any company (other Fisons UK) manufacturing gold or } gold/palladium ring targets for Biorad sputter-coaters? We have just been } informed (after waiting two weeks and having being informed that it was on } its way) that we have a 3-4 week wait for a new one from Fisons UK, a } slight problem as due to an "administrative oversight" there is no spare. } Any suggestions for an alternative source in future would be gratefully } received. } Yours faithfully } RE } } Richard Easingwood } Department of Anatomy and Structural Biology, } P.O. Box 913 } University of Otago, } Dunedin, New Zealand. } Fax:64-3-479 7254 } Telephone:64-3-479 7301 }
Message-ID: {MAILQUEUE-101.940621115228.256-at-eye1.eye.ufl.edu} To: microscopy-at-anlemc.msd.anl.gov
Several of our people are working with whole mounts of retinas removed from cow eyes, and are looking for a stain that will differentiate between dividing cells and inactive ones. We could use any suggestions for LM or fluorescent stains that do not require embedding and sectioning. Thanks! Evelyn Clausnitzer U of Florida Ophthalmolgy
Piotr Swierczynski was asking for specific stain for fibrocartilage. Don't think there is one-no-one here knows of one. Suggest you stain for collagen-thick bands in amongst cartilage would differentiate it. Possible stains would be Wright's, Masson-Trichrome or H and E/alcian blue double stain. Could also try an alizarin red S and Toluidine Blue O stain (Dawson 1926, Stain Techn. 1:123-4; Williams 1941, Stain Techn. 16:23-5). Let us know what you find out-which one works better.
Mark Elliott, UBC-Pulmonary Research Lab, St.Paul's Hospital Vancouver BC
Evelyn Clausnitzer was asking about stains for whole mounts for mitotic activity-try DAPI, Hoechst or acridine orange. They all work on tissue cultured cells which have not been embedded. Should be able to tell mitotic figures easily. Check with Molecular Probes to see if there are others, but these are fairly easy to use and look great.
Mark Elliott UBC-Pulmonary Research Lab St. Paul's Hospital Vancouver BC Canada
Have you tried just a basic H&E stain? It works well on fibrocartilage from a human intervertebral disk. Is the brain damaged? If so, have you tried: "Foot's Ammoniated Silver Carbonate Method"? From Foot (1924). It will show: vascular reticulum, tumor cells , and connective tissue around a brain tumor. Here is the procedure if you want to try it:
Fix: formalin (1:10) recommended; Cajal's FAB and Bouin's fluids also satisfactory.
Embed: in paraffin
Solutions: A. Ammonical silver carbonate: To 10 ml of 10.2% AgNO3 add conc. NH4OH, drop by drop until precipitate is almost redissolved; then add 10 ml of 3.1% Na2CO3, concentrated formalin, 1 ml; water, 100 ml. B. Reducing agent: 1% Na2CO3, 3 ml; conc. formalin, 1 ml; water, 100 ml. C. Intensifying solution: oxalic acid, 2 gm; conc. formalin, 1 ml; water, 100 ml.
Staining schedule: 1. remove paraffin from mounted sections and treat for 24 hr at room temp. in a mixture of pyridine and glycerol (2:1 by volume). 2. rinse in 95% etoh, then in water. 3. impregnate for 2.5 hr at 40 C in silver sol. A. 4. wash in distilled water. 5. reduce 5 min in sol. B. 6. wash in tap water. 7. tone 5 min in 0.2% gold chloride. 8. wash in tap water. 9. intensify by treating 5 min in sol. C. 10. rinse in tap water. 11. fix in 5-10% Na2S2O3.5H2O. 12. wash in tap water. 13. dehydrate. 14. cover in balsam or synthetic resin.
Note: The stain is not recommended for normal brain. It has considerable value in demonstrating connective tissue reaction around tumors if reticulin fibers are laid down by this connective tissue. If FAB fix is used, the reticulin staining is said to be suppresed.
ANNOUNCEMENT: Tribology and Coatings Technology Listserver/Mailreflector
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========================================================================== Nestor J. Zaluzec Materials Science Division , Bldg - 212 Argonne National Laboratory, Argonne, Ill. 60439 USA
Message-Id: {9406221358.AA27889-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb Subj: Re: Software / Font needed Orig-Author: {Brian Gregory Demczyk {temcom-at-engin.umich.edu} }:ddn:wpafb ----------------------------------------------------------- No one in their right mind would use Word Perfect instead of Microsoft Word! Of course one is able to overstrike in the latter!
Group - Per a recent request, the winners of the Polaraid International Instant Photomicrography Competition are: Grand Prize: Dr. Gerald T. Baker, Mississippi State Univ. B&/W Light Micrography: Floyd E. Alberts, Ford Motor Company Microscopy Techniques: John Georgiou, University of Toronto Material Sciences: Vito Giannini, Italcementi SpA (Italy) Student: Jim Wetzel: Clemson Univ. Color Light Micrography: Wutian Wu, Eastern Virginia Medical School Should any wish a copy of the full 3-page announcement, send me your fax number and I will be pleased to supply. I expect that we will do an article on the full winners list in a future issue of Microscopy Today. Regards, Don Grimes, Microscopy Today
Does anyone have a procedure for producing skeletal carbon TEM grids with holes sufficiently small to capture particles ranging in size from 200 nm to 30 nm?
Thanks.
-- Diandra L. Leslie-Pelecky Center for Materials Research and Analysis PH: (402) 472-9178 University of Nebraska FAX: (402) 472-2879 Lincoln, NE 68588-0113 diandra-at-unlinfo.unl.edu
I need to purchase an inverted epi microscope to conduct membrane kinetic studies. Because of the experimental setup I can't use immersion optics. I have about 22K to spend and I already have a good quality video camera. Are there any thoughts on which product lines are the best and recomentations for objectives?
} I need to purchase an inverted epi microscope to conduct membrane kinetic } studies. Because of the experimental setup I can't use immersion optics. I have } about 22K to spend and I already have a good quality video camera. Are there } any thoughts on which product lines are the best and recomentations for } objectives? } } Thanks } } Carlo Montemagno } } Try a Nikon, if you want a rotating stage a Nikon will not do. Clarissa
Subject: Time:2:46 PM OFFICE MEMO Re:Overbars in MS Word Date:6/22/94 It is possible to print nearly any selected character centered over nearly any other character by using, in combination, the 'Overstrike' and 'Superscript' typesetting commands provided in Microsoft Word. These commands are described in the section on producing formulas (which appears on pages 98 to 105 in the instruction manual for MSW V4.0). As an example, working with New York font, the following command string will produce a capital 'X' with a bar neatly centered above it: .\O.\AC(X,.\S.\UP11(_)), while the command string: (.\O.\AC(1,.\S.\UP11(_))0.\O.\AC(1,.\S.\UP11(_))) produces the set of Miller "bar one-zero-bar one". In these strings, the special character (.\) telling MSW to enter the typesetting mode is produced by holding down the command and option keys while typing a backslash (\). Command strings must be prepared by activating the "Show-#166#" mode , and the final printed format can be displayed by the "Hide-#166#" mode (obtained in the Edit menu or toggled by pressing the command and 'Y' keys simultaneously). Now, you may complain that it is inconvenient to type long command strings such as these as frequently as might be needed in many manuscripts. However, the process can be simplified by first typing the manuscript using a dummy variable (e.g. VX for vector X, 1B for overbar one, etc.). When completed in this manner, prepare the command string for a particular symbol and save it on the clipboard (command-c). Then use the Find utility (command-F) to find the first occurrence of the corresponding dummy variable, click in the ULH square to release the Find window, replace the dummy variable with the command string (command-V), find the next occurrence of the dummy variable (option-command-A), replace it, and so proceed through the entire manuscript. Try it, you'll find it works rather well. You will also find the full set of typesetting commands to be very useful for inserting formulas into manuscripts - they require a little practice, but turn out to be quite convenient and versatile, once you get familiar with the system.
Message-Id: {9406221911.AA29215-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb Subj: overstriking... Orig-Author: {"Daniel L. Callahan" {dlc-at-owlnet.rice.edu} }:ddn:wpafb -----------------------------------------------------------
If you find out how to overstrike in Word (Mac), please post to the microscopy listserver so all will know! I too have hunted this down unsuccessfully.
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu
Scott, To overstrike in Word for the Mac is staggeringly simple. To put e.g., a slash through a zero similar to one of the Scandinavian characters, you should i) hold down COMMAND and OPTION together and type a backslash ii) type an o (for overstrike strangely enough) iii)) type a LH bracket iv) type the two characters, separated by a comma v) close the brackets. (Of course the Scandinavian letters are available anyway so you would not bother doing this particular example) Congratulations, you have just overstruck! If you want to put a bar above an index, just make the second character a superscript hyphen.
Happy overstriking Rick Hall Mat.Sci., U. of Delaware
Okay folks, I'm more than a little suprised that Nestor hasn't chimed in here and I might be out of place, but this is _NOT_ the place to either discuss the pros and cons of a particular piece of word processing software, particularly when it has no direct bearing on microscopy, nor is this forum the place to flame someone for the signal to noise ratio of their posts. I have been the unfortunate participant (vicarious) in other flame wars on different forums and they serve _NO_ purpose. If you find the comments of someone offensive and wish to respond, please do so in private. Please, please, don't let this highly useful and informative discussion group dissolve into acrimonious shoot-outs over message content or perceived attitude. Thanks, I'll now fade back to my usual semi-transparent level of participation.
Dwight U. Beebe Institut de recherche en biologie vegetale beebed-at-ere.umontreal.ca Universite de Montreal Voice:514-872-4563 4101, rue Sherbrooke est FAX:514-872-9406 Montreal, PQ H1X 2B2 Canada
NB the command key is the key marked with an apple sign NB2 hold down the option, command and backslash (\) key simultaneously (this will activate the formula editor). NB3 it might be a bit longer then option-command-\O(1,_) but can be put in a macro
Cheers Timon
------------------------------------------------ Timon Fliervoet, Timonf-at-earth.ruu.nl, Faculty of Earth Sciences, Department of Geology, Utrecht University, P.O. Box 80.021 3508 TA Utrecht, the Netherlands. tel: (0)30 - 535054, fax: (0)30 - 537725
I just want to thank both Nestor & Dwight for bringing out the fact that this IS a microscopy list. I agree wholeheartedly that this is NOT the place for discussions (shall we say) on things such as word vs. wordperfect. Let's get back to microscopy please.
Just venting.
Thanks, Peling Melville
-------------------------------------------------------------- Peling Melville peling-at-amnh.org Interdepartmental Laboratories American Museum of Natural History
Reply_ Overstrike FONT Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu A company in Ann Arbor, Allotype Typographics sells two fonts that are great for Miller indices. They are called Haber and Thompson, they are based on Helvetica and Times respectively. I have been using these fonts for about 5 years now and have been very happy with them. Allotype can be reached at 313-480-3666. I am just a satisfied customer. Jfm.
We are trying to use a procedure that was established over 20 years ago. One of our problems is that a chemical was used to treat the glass slides in order to prevent the wood pulp fibres from sticking to them during drying. The chemical used was dichlorodimethylsilane which is considered highly toxic. We would like to hear any suggestions for an alternative way to prevent the fibres from sticking.
Greetings, I would like to say that the discussion on overstriking has been very helpful. While I agree that those parts of the discussion that "flamed" were not needed, the main discussion had a lot of content, and I don't see why this disscusion should not be allowed to continue on this group.
I bet word processing is the ONLY technique that EVERYone on this list uses. Word processing certainly is an indespensible technique for the professional microscopist. I guess it is reasonable to have some limits: certainly flames are unwanted, and so too discussions of an ethically dubious nature might be shunned, such as how microscopists can avoid income taxes, and finally discussions that are wholly irrelevant, such as on making the best cheese fondue could be canned. My view is that word processing is relevant for microscopists to talk about. I might add, as a light microsopist in a biology department, there are many subjects covered on the list that I ignore. Three cheers for clear subject lines.
To be fair to the individual who started the discussion thread on "Miller Indices" (not word processors!), the originator of the message, first sent the text to me and I was asked if it was appropriate for this forum. Since it was a serious question, and one which was related to microscopy and he was clearly in need of an answer (& and a listing of the various options for solving his problem) I concurred and told him to post the question to this forum. It's now run it's course and unless there is something else to add specifically on Miller Indices that has not been covered let's move on.
I will continue to be a behind the scene's filter for anything which the subscribers to this list may wish an opinion on prior to posting and a traffic cop when things appear to be getting shall we say "out of hand". Feel free to abuse my screen all you like. You will neither be the first and I'm sure you will not be the last, and as you are all aware the electronic trash can is only a button away.
I would have to agree with Tobias Baskin's comments on acceptable content of this bulletin board. As long as the material is something used by the readers on a professional level, why not include it as fair game? In an open system, such as this, it's up to the contributors to remain professional, and keep in mind what may be of general interest to the rest of the readers.
I find this forum interesting to read, and it's provided a wealth of information that I've squirreled away for future reference. I'd really like to see it kept open...
Thanks
Margaret E. Hogan, PhD Electron Microscopy Service The Jackson Laboratory 600 Main Street Bar Harbor, Maine 04609 (207) 288-3371 #1450 (207) 288-5079 FAX meh-at-aretha.jax.org
Message-ID: {MAILQUEUE-101.940623184732.288-at-UNDERBANK.shef.ac.uk} To: microscopy-at-anlemc.msd.anl.gov
Hi, I would have kept quiet but for some users suggesting that it is a good idea to exchange trivia about word processing. I joined a microscopy board, not a software panel - I get a lot of useless mail and I don't need a pile of messages about word processing. May I politely suggest that people who want to talk about anything except microscopy on this valuable service should form their own club.
As long as everyone is getting in the act, I will also jump in. I am a Mac user and I subscribe to a variety of newsgroups, many of which specialize in discussions of topics such as WP programs. I suggest that other Mac users seek out such discussion groups and PC users seek like discussions. Let's talk scatterin' events here...
Larry Sutter Michigan Technological University Department of Metallurgical & Materials Eng. 1400 Townsend Dr. Houghton, MI 49931
Does anyone have/know of a Photometrics Star 1 thermoelectrically-cooled CCD camera in good condition which is available for purchase? They are apparently no longer commercially available (that's why I am bothering this list about it.)
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Subject: Time: 4:42 PM OFFICE MEMO Is EM art? Date: 6/23/94 Are we scientists are artists? If the answer is the first then why are there so many micrograph competitions?
} Subject: Time: 4:42 PM } OFFICE MEMO Is EM art? Date: 6/23/94 } Are we scientists are artists? If the answer is the first then why are there } so many micrograph competitions?
} To answer this. EM is an art! You either have it or you don't!
} OFFICE MEMO Is EM art? Date: 6/23/94 } Are we scientists are artists? If the answer is the first then why are there } so many micrograph competitions? This question has no simple answer. For starters, the question of who (or what) an artist is may need to be answered. Answering the question, "What is a scientist?" is not easy, but at least science is a collection of fairly well pre/proscribed paradigms. Art, on the other hand, is a free-for-all. It is now generally accepted that photographs may be art; I say generally accepted because there are some very conservative/reactionary/retrograde critics who refuse photography art status, but if we relegate these people to the status of bonkers, then we'd all agree (as any reasonable person would) that photographs can be art. So what makes a micrograph, a specific class of photograph (although this definition must be broader to include video, computer representations, etc.) a work of art? Let's look at a more general question, what makes (an object) art? Or a question that attempts to seek boundaries: does art have to be made by an artist? If art can be defined by an audience's reception of an object or event and not by a performer (e.g. artist-- regardless of intent), then a micrograph can be a work of art without there being an artist, in this case a scientist. The danger this position raises, however, is that it may not priviledge only the relics of human endeavor; for instance, a rock or sunrise could be misconstrued as art. Here is the center of the issue: the objects to which the questions really refer (micrographs) are compelling aesthetic objects. Just because something looks good or touches that ineffable something in us and is made by a person or people does not mean it is art.
None of these musings lead directly to the question of why there are micrograph competitions. Entering a competition implies desire for exhibition or need for public recognition or motivation to WIN or the expectation of another line on the cv, but these apects of showing work, although the same or similar to concerns in the arts, really have nothing to do with art specifically.
The only way, logically, to answer the general question whether scientists are artists and to, pragmatically, avoid exploding the term "artist" so that it is meaningless, is to answer NO. More simply: There are scientists who are artists. Not all scientists are artists. The last two statements have nothing to do with whether micrographs, in general, are art. Some micrographs are art and some are not.
On Thu, 23 Jun 1994, rutledge phil wrote: } On 23 Jun 1994, Paul Webster wrote: } } OFFICE MEMO Is EM art? Date: 6/23/94 } } Are we scientists are artists? If the answer is the first then why are there } } so many micrograph competitions? } } To answer this. } EM is an art! You either have it or you don't! EM is a craft.
} Subject: Time: 4:42 PM } OFFICE MEMO Is EM art? Date: 6/23/94 } Are we scientists are artists? If the answer is the first then why are there } so many micrograph competitions?
From the for what its worth dept., since ya asked...:
I don't think that being a scientist should exclude one from appreciating art. Hopefully, its in the study of the symmetry and beauty of nature (be it biological or material, with a light or electron 'scope) that scientists can learn and "see" more of the world around them. Having such competitions, to myself anyway, allows one to view others' work; it gives a forum for the display of different photomicros that otherwise I might not ever see for myself. It also allows others to see the best of different techniques/methods.
Besides, its a way to win neato prizes :) :) But I suppose that ya have to enter 'em first, 'eh?
My {$0.02, -Rob ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /\v/\ | | Metallographic Lab. | Missouri Speleological Survey \-v-/ | | Rolla Research Center | Bat Conservation International /\v/\ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-rorc.usbm.gov | National Speleological Society #32993 /-v-\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
From HOLONET What's Appropriate? Please stick to microscopes! Reply to Tobias Baskin's message:
I was away from my office for one week and came back to over 150 E-mail messages from the Confocal newsgroup and from this newsgroup. It's going to take a long time to read and save the useful messages, and delete the redundant replies, and delete the all too large number of off-topic messages. Come to think of it, this reply is probably off-topic, so I would aprreciate your re-reading the Moderator's announcements of the purpose of this newsgroup instead of replying to this message.
} I would like to say that the discussion on overstriking has been very helpful...
I deleted all of those with extreme prejudice without reading any.
} I bet word processing is the ONLY technique that EVERYone on this list uses....
So what. I'm on here to learn about MICROSCOPES. I'll go as far as anti-vibration tables for acceptable hardware. Questions and comments on software not directly related to to your microscope should be asked elsewhere.
} there are many subjects covered on the list that I ignore...
Yes, but the junk mail still shows up in my mailbox.
} Three cheers for clear subject lines.
Agreed. I will delete without reading all future messages that have "None" or garbled subject lines (my apologies to any customers who don't get a reply from me because of this policy).
Message-Id: {MAILQUEUE-101.940624083209.352-at-bunyip.ph.rmit.edu.au} To: microscopy-at-anlemc.msd.anl.gov
I agree entirely John Millar RMIT Australia ------- Forwarded Message Follows ------- To: microscopy-at-anlemc.msd.anl.gov
Hi, I would have kept quiet but for some users suggesting that it is a good idea to exchange trivia about word processing. I joined a microscopy board, not a software panel - I get a lot of useless mail and I don't need a pile of messages about word processing. May I politely suggest that people who want to talk about anything except microscopy on this valuable service should form their own club.
Paul Fons asked about HREM image simulation software. In this context I would like to float an idea for responses. In the past, Hitachi has been selling our software at prices which (in my opinion) are ludicrously high. We are in the process of rewriting it for a RISC/UNIX workstation, exploiting X-windows and UNIX wide system calls. I am very tempted to make it freeware, either via ftpanon or otherwise when this is finished; in other words it should be compilable on any modern UNIX system (and PC/Mac systems as they move to be compatible with workstations).
As you know, it is a fallacy to imply that the two are mutually exclusive. EM is used as a tool in a great many areas of science. The yearning to use such a tool with some elegance (rather than in a manner that is dull or sloppy) is not unique to EM. I know biochemists, molecular biologists, and others, whose work has elegance. They are not satisfied with a mere dumping of required information (in any form) into a paper. They want the information to be well organized, tight, clear, a pleasure to read. One might say they do science with some art. I hope we can have more science involving EM that has this kind of elegance. If there is an occasional EM art show on the side, it is certainly not a problem.
Regards, Kent (A. Kent Christensen, Dept. of Anatomy and Cell Biology, Univ of Michigan, {akc-at-umich.edu} )
-----------------------------------
On 23 Jun 1994, Paul Webster wrote:
} Subject: Time: 4:42 PM } OFFICE MEMO Is EM art? Date: 6/23/94 } Are we scientists or artists? If the answer is the first then why are there } so many micrograph competitions? } } }
Reply_ RE} HREM simulation software Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu Absolutely excellent idea!!!!! There are too many people who write software in the microscopy field and think they are going to get rich on it. They try to sell it for exorbitant amounts of money and it is frequently buggy as all hell.They are usually unable to support it adequately since they are typically only selling it as a part time job. Software marketing and support is difficult to do properly! I think making it available free would be an excellent service to the user community.
-------------------------------------- Paul Fons asked about HREM image simulation software. In this context I would like to float an idea for responses. In the past, Hitachi has been selling our software at prices which (in my opinion) are ludicrously high. We are in the process of rewriting it for a RISC/UNIX workstation, exploiting X-windows and UNIX wide system calls. I am very tempted to make it freeware, either via ftpanon or otherwise when this is finished; in other words it should be compilable on any modern UNIX system (and PC/Mac systems as they move to be compatible with workstations).
Comments ?
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Why not? How can anyone be against this? If you can cover your costs somehow, and you want to do it, consider me a cheerleader!
On Fri, 24 Jun 1994, L. D. Marks wrote:
} Paul Fons asked about HREM image simulation software. In this } context I would like to float an idea for responses. In the past, } Hitachi has been selling our software at prices which (in my opinion) } are ludicrously high. We are in the process of rewriting it for a } RISC/UNIX workstation, exploiting X-windows and UNIX wide system calls. } I am very tempted to make it freeware, either via ftpanon or otherwise } when this is finished; in other words it should be compilable on } any modern UNIX system (and PC/Mac systems as they move to be } compatible with workstations). } } Comments ?
A synthetic Ni-olivine (Ni2SiO4) from the Smithsonian (USNM #717) may be available. We have some of this material that was used in a thermodynamic study by R. Robie et al (Amer. Mineralogist v.69, p.1096, 1984). You may be able to obtain some through Gene Jarosewich at the Smithsonian.
Jim McGee
James J. McGee email: jmcgee-at-lunatic.er.usgs.gov U.S. Geological Survey Phone: (703) 648-6782 959 National Center Fax: (703) 648-6789 Reston, VA 22092
Dear John, I assume the unknowns are silicate minerals. If not, you should make an attempt to have the same average Z-value for unknowns and standards--Chuck Fiori made a point of this in referrence to biological specimens. I'd try to mix a nickel compound (any one which is handy and doesn't have other peaks in the energy region of interest will do) with a molten silicate glass or with quartz if you can heat it enough to get thorough mixing. I'd make a range of different nickel fractions from ~1% to whatever you expect at maximum. The average Z and thorough mixing are the keys. Good luck.
As long as we are all voting, I agree that the discussion of word processors has run its course. However I did learn some useful things applicable not just to miller indexes but to microscopy writing in general. Since we are a varied background of users (and many of us could care less about subscribing to a bulletin board on word processors or operating systems, etc.) . IMHO the occasional inclusion of helpful but peripheral information and questions should be encouraged. Like others, my mailbox is too cluttered and I throw out a lot without reading. But I would like to keep things open. Occasionally a gem comes along.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Pathology * * Bowman Gray School of Medicine of Wake Forest University * * 910-716-4972 * * jjerome-at-isnet.is.wfu.edu * **************************************************************
RFC-822-Headers: Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII On Tue, 21 Jun 1994, Carlo Montemagno wrote:
} I need to purchase an inverted epi microscope to conduct membrane kinetic } studies. Because of the experimental setup I can't use immersion optics. I have } about 22K to spend and I already have a good quality video camera. Are there } any thoughts on which product lines are the best and recomentations for } objectives? } } Thanks } } Carlo Montemagno
On June 22nd Clarissa wrote in response to Carlo Montemagno's question:
} Try a Nikon, if you want a rotating stage a Nikon will not do.
For those who want a rotating stage for the Nikon inverted microscope, as of a few weeks ago, a rotating, centrable, gliding stage is available for the Nikon Diaphot 200/300. It attaches by the same 3 screws as the standard stages, it is not graduated, but works well for DIC. Cost: $1600.00 It is manufactured by Micro Video Instruments of Avon, Ma. USA For more information: 508 580 0080 or FAX 508 580 8623
I also heartily agree that open software would be a big advance. Don't limit it to HREM. The errors in commercial packages are virtually uncorrectable and we wind up with the situation bulldust in (the code) = bulldust out. Sadly only known after a lot of pain and wasted time. eg the K-line simulation problem tracked down by Alwyn Eades and his collaborators.
In another "hot" topic which could set off a mine is better than yours email war.....
JEC of GIT asked about imaging programs (and possible platform competition). Well let me try to be impartial and start the discussion ;-) ------------------------------------------------------------
There are many commerical programs available for handling image data, all of these do good to excellent jobs, (on both Mac's and PC's -- Oops just showed a preference didn't I--) unfortunately you will have to purchase them.
But to answer your question . No one has bothered to port NIH image to the PC platform. The lastest version 1.56beta9 runs on virtually all Mac's but not the PC. The source code is available on Zippy.nimh.nih.gov if you are interested in a challenge.
On the PC side the only freeware programs I am familiar with are NCSA Image for the PC, and UCFImage for the PC. There are also a couple of programs which I would call Viewers, the just let you put images up on the screen and not much else. All of these are freeware and will be available at the computer workshops at the PARIS ICEM13 meeting (July 17-22), as well as the at the New Orleans MSA/MAS meeting (August 1-5).
The NCSA suite of programs were not designed as an image processing programs but then again neither was NIH-Image, rather they might be better described as a data set visualization programs. I've occassionally used the NCSA code but not frequently, since I've found that I can more readily customize NIH Image to Microscopy related tasks.
UCFImage is a older image processsing type program out of the University of Central Florida, the version (5.0) I have lacks the polish of a true GUI program (it runs under DOS) and hence to most users who have been wedded to WINDOWS or Mac's (there I've now balanced my comments) might be disappointed. I found it clumbersome to use, however, if you are running on an older DOS machine then you may wish to check it out.
If anyone is still interested I can dig up the readme files and post segments tomorrow.
I'd be interesting in hearing about any other public domain programs which can be added to the software library database. So if you know of such please post the information here. as well as include the information on how to get copies. I'll then add the information to the EMMPDL libraries and try to insure that copies are available next month at both ICEM and MSA.
Received: From CORNELLC(MAL) by ANLNRH(ANJE6.10) for MICROSCO-at-ANLEMC; Sun, 26 Jun 94 22:49 Return-Path: {-at-CORNELLC.CIT.CORNELL.EDU:andy-at-EARWAX.PD.UWA.OZ.AU} Received: from CORNELLC (NJE origin SMT-at-CORNELLC) by CORNELLC.CIT.CORNELL.EDU (LMail V1.1d/1.7f) with BSMTP id 0700; Sun, 26 Jun 1994 23:47:52 -0400 Received: from uniwa.uwa.edu.au by CORNELLC.cit.cornell.edu (IBM VM SMTP V2R2) with TCP; Sun, 26 Jun 94 23:47:49 EDT Received: from earwax.pd.uwa.edu.au (earwax.pd.uwa.edu.au [130.95.156.3]) by uniwa.uwa.edu.au (8.6.8/8.6.4) with ESMTP id LAA22954 for {Microscopy-at-anlemc.BITNET} ; Mon, 27 Jun 1994 11:48:46 +0800 Received: from [130.95.124.120] (awsj [130.95.124.120]) by earwax.pd.uwa.edu.au (8.1C/8.1) with SMTP id LAA10562; Mon, 27 Jun 1994 11:47:40 +0800
I also heartily agree that open software would be a big advance. Don't limit it to HREM. The errors in commercial packages are virtually uncorrectable and we wind up with the situation bulldust in (the code) = bulldust out. Sadly only known after a lot of pain and wasted time. eg the K-line simulation problem tracked down by Alwyn Eades and his collaborators.
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I also heartily agree that open software would be a big advance. Don't limit it to HREM. The errors in commercial packages are virtually uncorrectable and we wind up with the situation bulldust in (the code) = bulldust out. Sadly only known after a lot of pain and wasted time. eg the K-line simulation problem tracked down by Alwyn Eades and his collaborators.
Anyone have a K-1905-1 board out of a Kevex micro-7000 EDX analyser spare that they are able to give away / sell? Ours is faulty and hindering one of our M.Sc. students.
Thankyou,
Keith Hallam
p.s. If I send a message out on the microscopy discussion group does it automatically get echoed to the usenet group too? If so, apologies for repeating myself!
We had a major network crash here at ANL over the weekend. FRIED fiber optics and lots of headaches all around. I apologize if messages got lost over the weekend. I think things are back to "semi-normal".
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I also heartily agree that open software would be a big advance. Don't limit it to HREM. The errors in commercial packages are virtually uncorrectable and we wind up with the situation bulldust in (the code) = bulldust out. Sadly only known after a lot of pain and wasted time. eg the K-line simulation problem tracked down by Alwyn Eades and his collaborators.
The image analysis facility down the hall is interested in the names of some software packages that a digitized or scanned electron diff pattern can be measured and indexed with. Are there any public domain/shareware programs available? Since the demand is small, they aren't willing to spend a great deal. Thanks in advance. Randy
___________________________________________________________________ Randy Nessler rnessler-at-emiris.iaf.uiowa.edu The University of Iowa Central Electron Microscopy Research Facility
Received: From CORNELLC(MAL) by ANLNRH(ANJE6.10) for MICROSCO-at-ANLEMC; Tue, 28 Jun 94 21:55 Return-Path: {-at-CORNELLC.CIT.CORNELL.EDU:paul-at-EARWAX.PD.UWA.EDU.AU} Received: from CORNELLC (NJE origin SMT-at-CORNELLC) by CORNELLC.CIT.CORNELL.EDU (LMail V1.1d/1.7f) with BSMTP id 2085; Tue, 28 Jun 1994 22:53:48 -0400 Received: from uniwa.uwa.edu.au by CORNELLC.cit.cornell.edu (IBM VM SMTP V2R2) with TCP; Tue, 28 Jun 94 22:53:46 EDT Received: from earwax.pd.uwa.edu.au (earwax.pd.uwa.edu.au [130.95.156.3]) by uniwa.uwa.edu.au (8.6.8/8.6.4) with ESMTP id KAA04414 for {Microscop-at-anlemc.BITNET} ; Wed, 29 Jun 1994 10:54:44 +0800 Received: from localhost (paul-at-localhost) by earwax.pd.uwa.edu.au (8.1C/8.1) id KAA19840; Wed, 29 Jun 1994 10:53:38 +0800
Why can't EM be both science and art - we use this tool as scientist and in the process create some wonderfully beautiful images that have a definite aesthetic value - in my opinion the fact that we can appreciate the beauty and have a desire to share it in no way detracts from our seriousness as scientists - besides it may grab the attention and interest of a non-scientist and get him or her interested in learning more about the subject of the image or the process by which it was created
JULY 1994 ISSUE OF THE JOURNAL OF MICROSCOPY - VOLUME 175 PART 1
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 1-9
Differential imaging in confocal microscopy
T. WILSON, R. JUSKAITIS & J. B. TAN, Department of Engineering, University of Oxford, Parks Road, Oxford, OX1 3QZ, U.K.
SUMMARY A coherent detection system using a two-mode optical fibre is described which permits confocal, differential amplitude contrast and differential phase contrast images to be obtained simultaneously from a scanning optical microscope. the direction of differentiation may be chosen arbitrarily. The differential imaging modes possess an optical sectioning property.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 10-20
A novel method of Z-contrast imaging in STEM applied to double labelling
W. TICHELAAR, C. FERGUSON, J.-C. OLIVO, K. R. LEONARD & M. HAIDER, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69012 Heidelberg, Germany
SUMMARY A novel method of Z-contrast imaging in the scanning transmission electron microscope (STEM) is presented. The technique relies on the element dependence of the angular distribution of the scattered electrons, and is realized with a detector consisting of a set of concentric rings. It is possible to discriminate 9-nm colloidal gold and silver specifically distributed on thin sections. In addition to this practical work, numerical evaluations are used to assess the method. With two smaller markers, this approach will be useful in discriminating closely-spaced antigenic sites when steric hindrance occurs with double-labelling using probes of different sizes.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 21-33
A new method to correlate acoustic spectroscopic microscopy (30MHz) and light microscopy
A. F. W. VAN DER STEEN, J. M. THIJSSEN, J. A. W. M. VAN DER LAAK, G. P. J. EBBEN & P. C. M. DE WILDE, University Hospital Nijmegen, PO Box 9101, 6500 HB Nijmegen, The Netherlands
SUMMARY A powerful new method is used to correlate between light microscopic and acoustic properties of biological tissues. Specimens of liver were sectioned into slices, between 250 and 10 micrometres thick. The thick sections were investigated acoustically, the thin sections by light microscopy. Markers that could be detected and located, both optically and acoustically, were used to find and reconstruct corresponding regions in the acoustic and optical sections (2.5 x 2.5 mm). Parameter images were reconstructed from the sections investigated acoustically. The acoustic parameters were attenuation at 30MHz, the slope of the attenuation spectrum (between 10 and 50MHz), backscattering at 30MHz, the slope of the backscattering spectrum (between 10 and 50MHz), and the local ultrasound velocity. Acoustic images were obtained in the frequency range from 10 to 50 MHz, yielding a lateral resolution of about 50 micrometres. The sections for light microscopy were stained according to the Goldner trichrome staining technique. The histological composition was determined quantitatively, using digital image segmentation techniques. The percentage of collagen-rich fibrous tissue, luminal structure and interstitial spaces, and the number of nuclei were calculated for regions of 250 x 250 micrometres. These histological features were correlated with acoustic parameters obtained from the corresponding regions in adjacent sections. It was thus possible to find the histological components responsible for acoustic parameters.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 34-43
High pressure freezing of cell suspensions in cellulose capillary tubes
H. HOHENBERG, K. MANNWEILER & M. MULLER*, *Laboratory for Electron Microscopy I, Institute of Cell Biology, Universitatsstrasse 2, CH-8092 Zurich, Switzerland. and Heinrich-Pette-Institute for Experimental Virology and Immunology, at the University of Hamburg, D-20251 Hamburg, Germany
SUMMARY A procedure for efficient cryoimmobilization of large volumes of cell suspensions or micro- organisms by high-pressure freezing is described. This procedure uses transparent, porous cellulose capillary tubes with an inner diameter of 200 micrometres, into which the suspensions are drawn by capillary action. The tubes are processed by high-pressure freezing and freeze substitution as if they were tissue samples. Centrifugation of suspensions at low temperatures is no longer necessary and cryopreparation is greatly facilitated. A very high yield of adequately frozen specimens is obtained due to the constant, defined sample geometry. This approach can also be used to process suspensions by conventional chemical fixation, eliminating the need to embed pellets in low-melting-point agarose, for example, prior to chemical fixation. The preparation procedure is demonstrated with suspensions of nematodes, paramecia and bacteria.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 44-53
Nucleotide and protein distribution in BrdU-labelled polytene chromosomes revealed by ion probe mass spectrometry
RICCARDO LEVI-SETTI, JAN M CHABALA & SARAH SMOLIK*, Enrico Fermi Institute, The University of Chicago, 5640 South Ellis Avenue, Chicago IL 60637, U.S.A., and *Vollum Institute for Advanced Medical Research, Oregon Health Services University, Portland, OR 97201, U.S.A.
SUMMARY Detailed chemical maps of BrdU-labelled polytene chromosomes of Drosophila melanogaster, obtained by imaging secondary ion mass spectrometry, reveal separately the distribution of DNA and proteins in the chromosomes. The thymidine-analogue BrdU within the chromosomal DNA is localized by detecting the Br- secondary ion signal, while both nucleic acid and protein content, are mapped through the abundantly emitted CN- signal. This novel approach supersedes, and helps to explain the origin of, the banding patterns that are observed by conventional staining techniques. the high spatial resolution and chemical and isotropic sensitivity of the technique should enhance the localization of specific genes by in situ hybridization in mitotic chromosomes.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 54-59
Application of backscattered electron imaging to the study of the contact zone between lichen and rock. Assessment preparative procedures
J. WIERZCHOS & C. ASCASO, Centro de Ciencias Medioambientales, Serrano 115, bis Madrid 28006, Spain
SUMMARY In the study of the lichen-rock interface, light microscopy, scanning electron microscopy (SEM) in secondary emission mode and transmission electron microscopy are the most commonly-used techniques. As these methods have some limitations, there is a need to explore other techniques for observation of the lichen-substrate interface. One of the most promising methods is the application of SEM in the back-scattered electron (BSE) emission mode. The thallus of Aspicilia intermutans (Nyl.) Arn. growing on granitic rock was examined by SEM in BSE mode. The detailed preparation of transverse sections of the lichen-rock contact zone is presented. The BSE scanning images of the lichen-rock interface obtained present new insights into the ultrastructural features of the biological components, providing more information about the biogeophysical and biogeochemical weathering of rock.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 60-69
Semi-automated measurement of true chord length distributions and moments by video microscopy and image analysis
EBEN H. OLDMIXON, JAMES P. BUTLER* & FREDERIC G. HOPPIN, Division of Pulmonary Medicine, Memorial Hospital of Rhode Island, 111 Brewster Street, Pawtucket, RI 02860, U.S.A. and *Harvard School of Public Health, Boston MA 02115-9957, U.S.A.
SUMMARY The distribution of lengths of airspace chords in pulmonary parenchyma characterizes many architectural features of the alveoli and alveolar ducts. Laborious to obtain manually, the distributions and density functions may be acquired semi-automatically by video microscopy, digitization and image processing. The accuracy of the estimation is influenced by the microscopical methods and also by the techniques used (i) to convert the digitized grey-scale picture to a two-valued image, (ii) to collect the chord lengths and (iii) to compensate for finite field widths. The last problem arises because some of the chords are completely visible within a field, while others are only partially seen, since one of the two air-space boundaries lies outside the field of view. The error systematically biases the observed distribution. This paper presents solutions to hardware, software and analytic problems encountered while developing the capability to measure airspace chord length density functions semi- automatically. Formulas for estimating the true chord length density function from samples of observed chord lengths are presented. Also given are formulas for the estimation of the first and second moments of the true chord length distribution from the means of observed chord lengths. These techniques of image preparation and analysis should be suitable for characterizing particle, grain or cell size distributions, especially where many profiles fall partially outside the field of view.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 70-83
A simple algorithm to measure the volume-weighted and number-weighted mean volume of particles
GUY J. LAROYE & K. GRANT, Ontario Cancer Institute, 500 Sherbourne Street, Toronto, Ontario M4X 1K9, Canada
SUMMARY An algorithm is presented which offers an alternative approach for measuring volume- and number-weighted mean volume and standard deviation of particles. Using a computer- assisted manual method the following intermediate steps are performed automatically: generation of linear probes emanating from the sampling point of the object and intersecting the profile periphery, measurement of their lengths, and measurement of the area of the transect required for estimating the standard deviation of the volume-weighted mean volume. By first tracing manually the outline of the periphery of the object with a cursor, on a magnetic tablet or on an image acquired into the computer with a video camera, the location of all pixels of the periphery is registered and the area of the transect is measured concurrently. The computer is informed of the coordinates of the selection point in the uniform random (UR) sampling grid by clicking the cursor. All ensuing random linear probes between the sampling point and the object profile periphery emanating from this selection point, radiating at angular intervals of 29-30 degrees to the periphery. In the case of vertical sections, similar lines are generated at intervals where the sine of the angle changes by a value of 0.33. The volume-weighted mean volume of the object is estimated from the average of all products. As the periphery is traced, the algorithm can automatically determine the area of the cross-section of the object, from which the standard deviation of the volume-weighted mean volume can be calculated. Some elements of the algorithm are also used for the measurement of the number-weighted mean volume. The latter procedure is facilitated using an acoustic vertical depth monitor attached to the microscope. The impact of truncation ('lost caps') on the precision of measurements is discussed. The algorithm is of particular use in light microscopy for measuring cell nuclei by direct visual inspection of the microscopic field using a side-arm mirror assembly interfaced with a magnetic tablet.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 84-89
Microspectroscopic measurement of optical properties of rat liver in the visible region
AKITOSHI SEIYAMA, SHENG-SONG CHEN, HIROAKI KOSAKA & TAKESHI SHIGA, Department of Physiology, Osaka University Medical School, 2-2 Yamadaoka, Suita, Osaka 565, Japan
SUMMARY Microspectroscopy is used to investigate optical properties of haemoglobin-free perfused rat liver. Visible spectra of 20 micrometre diameter spot size were measured in transmission and/or reflection modes as a function of the thickness ( {1200 micrometres) of the liver edge. Optical density (OD) in transmission mode increased with increasing liver thickness, whereas in reflection mode OD decreased but became almost constant above a certain thickness (c.600 micrometres) of the liver. The Kubelka-Munk (KM) two-flux model, with a minor modification, was applied successfully to the analysis of the changes in OD as a function of the thickness. This approach estimates the KM absorption coefficient, KM scattering coefficient and effective penetration depth of the liver. The optical properties were similar to reported values, obtained with different methods.
Journal of Microscopy, Vol. 175, Pt. 1, July 1994, pp. 90
Letter to the Editors: A generalized terminology for multidimensional microscopy
MARK A BROWNE, C VYVYAN HOWARD, GLEN JOLLEYS & DUNCAN STACEY, Department of Fetal & Infant Pathology, University of Liverpool, PO Box 147, Liverpool, L69 3BX,
Gillian Wilson Executive Editor The Royal Microscopical Society Journal of Microscopy & 37/38 St Clements Proceedings of the RMS Oxford OX4 1AJ UNITED KINGDOM rms-at-uk.ac.ox.vax OR rms-at-vax.ox.ac.uk Telephone +44 (0)865 248768 Fax +44 (0)865 791237
Comments: Converted from PROFS to RFC822 format by PUMP V2.2X
} ALSO...we have an OMU-3 which we LIKE to repair, but I was told some time } ago that parts for it are also no longer available. Is anyone aware of } a parts inventory and/or service for this instrument. }
Try calling TEK-NET, Inc. I haven't used them, but spoke with someone there recently, so I'm pretty sure they're still in business. Their letter of introduction, sent out in '90, described the organization as a group of service engineers who used to work at Cambridge Instruments' East Region Service Center, until that center was closed. The list of equipment that they service is impressive, and Reichert is among them.
I've been tempted to post this information before, just to let folks know they exist, so here it is:
TEK-NET, Inc. 1985 Swarthmore Ave. Lakewood, New Jersey 08701 (908) 905-5530 (908) 905-0152 FAX
I have no affiliation with them, just wanted to get the word out.
John chandler-at-lamar.ColoState.EDU Fort Collins CO
This fall we are starting a new graduate level course on all aspects of light microscopy ie. fluorscence(sp),polorized,bright/dark field etc. and need recommendations for a good textbook. This course will include epquiment and methods. Thanks in advance for any help.