I don't ever remember having spoken to you regarding Visilog. By the way, Optimas does not generate C code in their recorder that's why they use the expression C like, actually it is a language called ALI which is not compatible to Visual C++ in any way.
You say you don't have any interest in promoting their product but you sure sound like it.
For your information, if you had looked at Visilog you would have noticed the following features:
- much wider selection of imaging algorithms than other Windows based sofware.
- wide selection of frame grabbers, Matrox Meteor, Pulsar and now including support for the new Matrox Genesis board using the C80. ALso COreco's TCI, Integral Flashpoint and ITI IC PCI plus new drivers coming for Data Translation and EDT.
- support for NT, Win95 and all Unix workstations.
- Real time morphology using the New Matrox Genesis C80. Pentium Pro's using WindowsNT do not come close to the power of this processor. No other off the shelf software supports dsp based processors like Visilog does.
- The ability to process floating point images for images which are greater than 8 bits per pixel.
- a powerfull c interpreter which generates real C code compatible with Visual Basic and VIsual C++.
- C interpreter for generating low level code and creating stand alone applications.
- Outstanding support and customer service.
- Run time version for as low as 1000.00
- 3 versions of the software starting at 1,000.00$
- On site training and consulting.
- A big and loyal installation base in the US, Europe and Japan,
- new easy to use user interface.
By the way, we will be at the Microscopy show in Cleveland, stop by and I'll give you a good demo. You can also see our new 3d Reconstruction package running on Windows NT.
Regards,
At 02:32 PM 7/31/97 -0400, David_Bell-at-Millipore.com wrote:
} I was wondering if anyone has worked out a formula on whether to charge per } sample or per hour.
It depends on what the lab does.
From the point of view of your "customers" the per sample rates are preferable because then their costs are predictable. It's also easier for you to do the bookkeeping, since all you have to do is count the samples.
On the other hand, if you do non-routine work, there is no way you can do a per sample pricing and come out fair.
As we do a mixture of routine and non-routine stuff, we have a mixed price system. For the routine analyses, we have a per sample rate based on the average time it takes us to handle the sample. In our catalog, we have specified EXACTLY what is included in these routines. Anything that differs from these standard procedures at all, goes on the hourly rate. When we apply the hourly rate, our customers (all internal) have the opportunity of specifying an upper threshold value. When we see that the analysis is going to be more expensive than this value, we call them up and ask them if we should continue. If they say no, then we charge them for the time we spent and give them the results obtained so far (if any).
Hello All, Has any-one any experience with Boehringer Mannheim's antibody to GABAA Receptor alpha chain, specifically whether it cross reacts with rat alpha chain?
Does anyone know of any other available antibodies to GABAA Receptor alpha subunits? I've checked all the usual commercial sources I could think of, with no luck, and would appreciate any suggestions.
Thanks Sharon
Dr. Sharon Miksys Department of Pharmacology University of Toronto 1 King's College Circle Toronto, Ontario Canada, M5S 1A8 Tel: (416) 978-4082 Fax: (416) 978-6395 Email: s.miksys-at-utoronto.ca
For what its worth, I share many of the same concerns with respect to systems that require proprietary hardware. There are many IA systems that will perform a variety of tasks. There are also a wide range of levels of sophistication in many of the IA systems offered to date. I suppose the best choice is determined by the type of work you will be doing, i.e. if you will routinely perform a similar task day in and day out, then a turn key system is probably the best bet, otherwise, you need a system with versatility, and ease of use, key word being EASE OF USE.
In general most IA systems all offer a platform of similar operations, upon these, are added some features that make their software "different" from the pack. Many IA systems have a sleek, and hi tech look, but behind the scenes they are doing many of the same operations as the next IA system. To me the key point (assuming the system is full featured of course) of any IA system is ease of use, ease of programming, ease of program modification
If you are going to be faced with a wide variety of applications, select a system that will be able to handle sample variation during analysis, and that you will intuitively be able to run without spending months trying to learn a programming language. Most of us are not programmers, therefore, ease of programming is paramount. Most systems I have seen allow you to write a sophisticated macros, but they often do not easily allow you to fine tune, or tweak your program without having a good working knowledge of their programming language. In this light, I feel a good deal of attention should be placed on how easy it is to write and modify macros.
Two systems to date that I believe are the easiest to program and modify: On the high end, Kontron KS400, and on the low end Mediacybernetics Image Pro Plus. These two IA programs offer a broad spectrum of applications, and have a good support team.
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Tony:
Some principles I think are important. Stay away from systems that require proprietary hardware boards for the software to run. These systems tend to become obsolete quickly or are expensive to upgrade. The less expensive image analysis software programs tend to be easy to use but lack the flexibility and power when confronted with a difficult problem. For overall cost and performance, I think PC based systems are the best. My lab has chosen Optimas software (runs under Win95 or NT) as the main image analysis platform, and sofar it has been able to do everthing we require.
Regards,
John J. Turek, Ph.D. Associate Professor Director, Electron Microscopy Laboratory and Core Laboratory for Image Analysis and Multidimensional Applications (CRISTAL) Department of Basic Medical Sciences 1246 Lynn Hall, G193C Purdue University W. Lafayette, IN 47907-1246 Phone: 765-494-5854 Fax: 765-494-0781 Email: jjt-at-vet.purdue.edu
No, we never did speak regarding Visilog. This may be due to the fact that I do not live in Canada, but even if you are the sales rep for the Northeast, we tried to keep sales people out of the examination, and talk to end users. Only after we had narrowed the choices down to what we thought would meet our needs did we bring the sales people into the picture. I do not believe that in my original memo, I stated anything negative regarding Visilog, but the fact of the matter is, we found that several people considered it not very user friendly (this may have been an older version). With regards to the ALI language not being true C, this is true, but most anything that can be accomplished in C can be done with ALI (maybe it won't be as elegant a program, but it will work). The reality is that ALI is a vector based language and the reason we are doing image analysis is to crunch numbers, and the most efficient way to handle large amounts of numbers is with vectors. As to the rest of your "unique features", it seems that Optimas meets most of them with the possible exceptions of the Matrox Genesis C80 and the price. Oh, by the way, your starting price may be $1000.00, but what does the average system go for?
With regards to your inference that I have an interest in promoting Optimas, I am not on any payroll of any organization that promotes any software, and I do not appreciate your implying that I am. I thought that the purpose of this list serve was for people to give their opinions on a subject when they have one, and that is what I did. I do not think that this is the forum for sales people to make judgments on other people's opinions and I would be interested in hearing what other microscopy scientists and professionals feel about this.
Regards,
David
ln-at-noesisvision.com on 08/01/97 02:11:28 AM
To: David Bell, bruton-at-EMU.UNP.AC.ZA cc: Microscopy-at-sparc5.microscopy.com
The Noesis office is in Canada, just like Matrox, Coreco and Dipix.
But we sell in the United States mainly through a wide number of dealers in every state and region.
Just like thousands of US corporations are located in the US and sell directly to Canada. The opposite also exists.
I can assure you Visilog provides a much wider selection of features than Optimas with our high end version at 6,000.00. Our 3,000.00$ version is similar to Optimas.
Thanks anyway.
At 11:16 AM 8/1/97 -0400, David_Bell-at-millipore.com wrote:
} Dear Luc Nocente,
}
} No, we never did speak regarding Visilog. This may be due to the fact that
} I do not live in Canada, but even if you are the sales rep for the
} Northeast, we tried to keep sales people out of the examination, and talk
} to end users. Only after we had narrowed the choices down to what we
} thought would meet our needs did we bring the sales people into the
} picture. I do not believe that in my original memo, I stated anything
} negative regarding Visilog, but the fact of the matter is, we found that
} several people considered it not very user friendly (this may have been an
} older version). With regards to the ALI language not being true C, this is
} true, but most anything that can be accomplished in C can be done with ALI
} (maybe it won't be as elegant a program, but it will work). The reality is
} that ALI is a vector based language and the reason we are doing image
} analysis is to crunch numbers, and the most efficient way to handle large
} amounts of numbers is with vectors. As to the rest of your "unique
} features", it seems that Optimas meets most of them with the possible
} exceptions of the Matrox Genesis C80 and the price. Oh, by the way, your
} starting price may be $1000.00, but what does the average system go for?
}
} With regards to your inference that I have an interest in promoting
} Optimas, I am not on any payroll of any organization that promotes any
} software, and I do not appreciate your implying that I am. I thought that
} the purpose of this list serve was for people to give their opinions on a
} subject when they have one, and that is what I did. I do not think that
} this is the forum for sales people to make judgments on other people's
} opinions and I would be interested in hearing what other microscopy
Course Announcement: "Optimizing Light Microscopy" When/Where: (a) New York City, November 3,1997 (b) Springfield, MA November 5, 1997 (c) Boston, MA November 7,1997 What: a lively, fast-paced slide lecture and demonstraton for anyone how uses or plans to use a light microscope: students, teachers, medical technologists, clinicians, pathologists, and lab managers. Beginning to more experienced practictioners welcome.
For details... (a) read below (b) send for brochure (c) visit the Microscopy/Microscopy Education booth at MSA - #502
Program: 1. A quick tour around the microscope - getting to know the bits & pieces 2. Koehler illumination & you: 4 critical steps for aligning you and your microscope to reduce headaches, fatigue, and errors 3. Care and cleaning 4. Useful principles for understanding and optimizing imaging 5. Putting the basics to work: a. Troubleshooting b. Understanding Phase and Hoffman Modulation Contrast 6. The Video connection: cameras, computers, and your microscope 7. Bringing out the best: quick, easy, and often free techniques for improving contrast 8. Advanced contrast techniques: Fluorescence and DIC 9. Becoming a better consumer: matching your microscope to your application 10. Questions, Answers, and Information Exchange (Note: Instructor may vary class content slightly to meet the needs of participants)
Free with your tuition: "Optimizing Light Microscopy for Biological and Clinical Laboratories" (Kendall-Hunt, 1997). Nearly 200 pages of helpful hints, quick experiments, and new iedas for getting the best from your microscope. Hot off the presses!
CEU's: 0.6 CEUs, 6 P.A.C.E CEU's Microscopy/Microscopy Education adheres to the guidelines established by the IACET.
Pricing: $150 (includes tuition, breaks, course materials, and copy of book) *****Save $25 if paid by 10/17/97.***** Send three from the same facility and save $50 on tuition for the third person.
Refund policy: Full refund for cancellations made by 10/17/97. After that date, 50% refund or full credit for future class. Substitutions accepted.
Questions: call Barbara Foster or Dr. Ken Piel at MME: (413)746-6931
Registration: Download the form below and fax to (413)746-9311 or call (413)746-6931 and ask for Ken.
Check course you will be attending: ___ New York City, November 3 (#971103) ___ Springfield, November 5 (#971105)* ___ Boston, November 7 (#971107)
Method of Payment: ____ Check enclosed for $ _______________ ____ Visa ____ Mastercard Name on credit card: __________________________________ Credit card number: ___________________________________ Expiration date: ______________________________________ ***If billing address is different from one shown above, please show billing address below: _______________________________________________________ _______________________________________________________ _______________________________________________________
Stephen A. Shaffer wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Inter/Micro 97 Day 4 } } I'll start with some notes on Wednesday's sessions since I neglected to } summarize them last night. I was, er, a bit slow by the time I got } back. :-P The session focus on Wednesday was "History and Art," } although that was only loosely held to. Several of the papers dealt } with artistic subjects but not art conservation per se. } } Gary Laughlin spoke about "A Unique Metallurgical Process From the Early } Bronze Age" in which he described the findings at, and significance of, } a site excavated at the Kestel Mine in the Taurus Mountains of Turkey. } The site dates from the third millenium BC. The examinations indicate } that cassiterite ore was mined and refined at the site to yield a black } magnetic oxide. This would have been more readily separated, due to its } magnetism, than could be accomplished by other means. } } Dr. McCrone gave an "Update on the Turin Shroud" in which he reviewed } several letters he received from Father Rinaldi prior to the Father's } death in 1993. In the letters, Father Rinaldi effectively acknowledged } the proof that the Shroud was painted and actually dated from much later } than the time of Christ's crucifiction, thus was not the true article. } } (For those of you who may not know, Dr. McCrone concluded early on in } the Shroud investigations, and from microscopic observations alone, that } the shroud was a painting. He stood nearly alone in this view and was } vilified for nearly two decades before Carbon dating ultimately } confirmed his conclusions.) } } John Delly gave one of his typical, amazing presentations, this one on } "Hand-colored Microscopical Illustrations." In it John took his } audience on a delightful stroll through 19th century microscopy } publications illuminated with hand-colored illustrations. He } demonstrated the evolution and later de-evolution of the quality of such } illustrations, the variation that one can see from different } illustrators of the same work, and the differences that are seen } edition-to-edition of the same work. Of course, when he became } interested in the subject, John felt compelled to master the art } himself. Through his own study and practice he gained the insignt } necessary to understand and explain the techniques and variations seen } in these historical works. The illustrations are, indeed, beautiful and } many of us are fortunate to own examples of these illustrations in early } works on microscopy. Because of the vast number of color illustrations } necessary to address his subject, it is unlikely that this presentation } will ever be recorded fully in print. (How about a book, John?) Those } of us fortunate enough to be in the room may be the only ones ever to } enjoy this particular product of John's efforts. Thank you, John, once } again. } } Have you ever stood befor an audience wondering why on earth you found } yourself presenting in the particular session where you were? Wayne } Moorehead must have when he spoke on "A Tale of Two Danas; Influences in } Mineralogy" but he soldiered on and did a fine job chronicaling the } lives of two remarkable men. The mineralogists in the audience will } need no introduction to the Danas but I'll just mention for the others } that the elder Dana published his first edition of the System of } Mineralogy in 1835 at the tender age of 24. It was the first such major } scientific work of classification written in English and he and his son } went on to publish or edit a vast array of classic works in Mineralogy. } Together or individually, they edited the prestigious American Journal } of Science continuously for an astounding 95 years, from 1840 to 1935. } One of the most noteable achievements that Wayne mentioned, in my mind } anyway, was when James Dana, in the introduction to a revised edition of } his classic System, abruptly abandoned his entire earlier classification } system as outdated!. Believing that system no longer consistant with } emerging thought, he just as abruptly adopted and described a newer } system which largely stays with us today. I find such honest } self-appraisal and the ability to continue to move forward without } missing stride quite refreshing. } } Wednesday afternoon was occupied by two sessions which would be unusual } at other meetings. Using video microscopy, Anna Teetsov of McCrone } Associates demonstrated some micromanipulation techniques within the } context of creating artistic works on microscope slides by arranging } butterfly scales of various colors into micro-images. Anna and a few } others continue to develop this art form which is particulary unique to } the community of microscopists. One has to have a microscope and } micro-related knowledge and skills in order to produce these beautiful } little creations, then one has to have a microscope to view them as } well. Kind of nice, don't you think? Something we can hold purely for } the aesthetic pleasure and uniquely our own. } } The afternoon was closed with a demonstration by Alan Shin on how one } can construct a working replica of Leeuwenhoek's single lens } microscopes. I did not attend this demonstration as I have on a } previous occasion taken a longer version from Alan in which we got to } actually construct our own microscopes. Comments from those who did } attend and look through the instrument Alan made reflected surprise at } how much one could see and pleasure at the experience of seeing an } insturment of such historical significance actually fabricated. } } Today's sessions were on Forensic Microscopy. Jose Almirall told us } about "Developments in Glass Examination: Automated Microscopy } Techniques and Composition Analysis." Jose's talk was very interesting } and perhaps somewhat troubling to practicing forensic scientists as he } told us of (among other things) a remarkable consistancy in the optical } properties of some glasses, especially window glass manufactured by the } "float" process. The new information for me was the time over which the } products of these plants will remain indistinguishable under } conventional forensic examination techniques. I am not aware of other } time-dependant studies of glass properties but Jose showed data } collected over at least 18 months, during which the product of a float } glass plant was entirely uniform in refractive index. He did however, } offer a remedy for this disturbing finding. He showed that glass } samples which are indistinguishable by refractive index can often be } distinguished by elemental analysis of Fe, Mg, Al, and Zr using } ICP/AES. Now all the forensic people have to do is get themselves one } of these and... ;-) } } Wayne Moorehead gave another excellent paper on Thursday, this one on } "An Introduction to Microscopical Feather Identification." Wayne told } us that the flight and tail feathers of birds are not always useful for } identification but that the down or contour (breast) feathers can be } distinctive, at least down to the order of birds, occasionally to the } family, but virtually never to the genus or species. Still, } identification at this level may prove very useful in a forensic case. } Wayne illustrated how appropriate preparations can be made, what } features of the feather barbule to examine and how they can vary. He } also showed and described the identifying characteristic of numerous } feather types. } } Thom Hopen gave an interesting talk on "Teaching Forensic Microscopy in } Countries Formerly Known as the Soviet Union." Thom responded to a } State Department request that he make numerous trips to various } countries of the former Soviet Union. He has taught courses of fiber } and paint comparison, explosives residue analysis, and basic } microscopy. He found his students to be highly motivated and dedicated } people, anxious for quality instruction in basic forensic microscopy } techniques. Often they are at least adequately equiped though sometimes } have little or no idea how to fully exploit the equipment they have. } (Unfortunately, when it comes to microscopy, this is too often true here } also! My comment, not Thom's.) One can only immagine the difficulty of } teaching in a completely and literally foreign environment, working } through a translator, and using instrumentation previously never seen. } Often, Thom had to set the equipment in proper working order prior to } beginning instruction. But apparently all has worked out for him and } his students and several more trips are planned to continue the } education. } } Well folks, I think I'll stop there. Of course, there were many more } fine presentations and, once again, I'll mention that my choice of } topics covered here in no way reflects badly on the other papers. All } of the presentations were excellent. } } Tomorrow is given over to a tutorial workshop on the Dispersion Staining } technique. It will be given at McCrone Research Institute by Dr. } McCrone and will be attended by twenty-odd students, all that can } reasonably be accomodated in such a hands-on session. For the rest of } us, this afternoon marked the end of another educational, enlightening, } and just plain fun Inter/Micro. } } Special thanks, as usual, to Nancy Daerr who coordinates all } arrangements for these meetings and who, as usual, did an exceptional } job of taking care of us and making our stay wholly enjoyable. } } To all of those interested in these meetings, please note: Next year } marks the Golden Anniversary of Inter/Micro, the fiftieth anniversary. } (Wow!) Plan on attending what promises to be an excellent meeting. } Contact Nancy Daerr for further information, to be put on a mailing } list, etc. She can be reached at McCrone Research Institute, 2820 S. } Michigan Avenue, Chicago, IL, 60616 or simply as ndaerr-at-mcri.org. The } phone numbers at McRI are 312-842-7100 (voice) and 312-842-1078 (fax). } } It's been a pleasure being your ears at Inter/Micro 97. But } tomorrow... Ahhh, Chicago! The architecture, the museums, the Art } Institute! I feel like a nice walk! Happy trails to all, and to all, } Good Night. } } Steve Shaffer } MicroDataware } sshaffer-at-microdataware.comDear Steve,
Many thanks for keeping us all up to date on this valuable meeting. Summaries from one day would have been really nice but summaries from all four were a gift. They were much appreciated.
I am afraid like many many other SEM operators you are using far too high= a kV. If you operate a SEM at 15kV plus you rarely see the true surface of=
the specimen under normal observation conditions. The manufacturers have=
cottoned on to this and improved the lower kV performance tremendously ov= er the last 15 years. You may have noticed how the maximum kV offered was 4= 0 or even 60kV in the early 70's whilst now many offer only 25kV. I rarely=
use a SEM above 10kV unless I am after more sub surface detail when I too=
will use up to 30kV, but only for this reason!
Your problems:-
1. You are using 15kV plus, much much too high with fragile or sensitive specimens. CURE - come down to {7kV 2 to 5 would be best if the sample is really sensitive.
2, You find at lower kV that you simply do not generate enough signa= l to make operation possible at the resolution that you require. =
CURE - move the filament forward in the cathode until you can obtain at least 30uA emission at saturation with the bias set to give th= e highest emission. Without this level of signal sure your task will be ve= ry difficult.
3. The system lacks signal and performance when you lower the kV. CURE - lift the sample in the system because lowering the kV will=
have increased the lens aberrations. Lifting the sample nearer to the le= ns will reduce the aberrations and in doing so the same number of electrons = as you have used at a lower WD will be better packaged giving you a higher current density and a higher signal. How high is high you may ask? Well= I would not dream of operating at {7kV with a WD greater than 10mm, with 3 = to 5 mm being my target depending on the make of instrument. I do not know the instrument you have intimately but if it has a conical lens 3 to 5 mm=
is fine, if it has the old fashioned big flat bottomed lens then 5 to 7mm=
may be better.
4. Resolution is difficult to specify but at 3kV I would expect a correctly set up electron gun to enable you to work with W at 15,000X. I= f Amray offer a low kV anode this would help a great deal. An alternative = is to lift the anode by fitting spacers underneath it! The normal anode to cathode distance is about 1mm for every 2kV. If you are able to lift the=
anode by about 5mm this would make a great deal fo difference to the gun performance. WARNING - PLACE A SIGN ON THE INSTRUMENT "NOT TO BE RUN ABOVE 5kV" whilst you are using it and remove the anode modification prio= r to leaving the instrument.
5. Filament life is going to come into this performance equation. I=
hate people who boast how long their filaments last, I liken this to leaving my car at home whilst I am abroad as I find this to be the most economical use of my car, it doesn't seem to use any fuel at all :-) If you really use a filament it will not last very long but if it makes the impossible possible who cares???
Hope this helps please come back if you need more.
I used to do platelets when I first started in this business. I centrifuged the whole blood (in citrated tube) for a few minutes in a tabletop centrifuge to separate the buffy coat. I then carefully dropped my standard fixative (2+2 glutaraldehyde/paraformaldehyde in 0.1M phosphate buffer) into the tube. After a few hours the buffy coat containing the platelets was lifted out like a pill which could be razor cut in pie-shaped slices. Those were then osmicated, dehydrated, and embedded like any other tissue. Joyce Craig Chicago State University
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Please unsuscribe me from the list. Thanks!
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Continuing the discussion of this topic. I used twin jet electropolishing a bit about 1970, but was frustrated by not being able to determine when a shiny surface was produced. (I had been able to see the sample in an old glass system operated manually). About that time, one of the early South Bay 550 polishers was ordered by me here at Argonne because it permitted magnified, IN SITU, viewing of the sample during polishing. This was needed to thin 1 or 2 new materials every week! The ease of use permitted accumulating reproducible data published in a 65 page report, (ANL-80-120), used world wide, a journal cover photo, and 12 other published articles. This work brought me the MSA "Technologist of the Year" award in 1994. About six 550 polishers are used exclusively at Argonne-some as long as 25 years! The unit can do jetting from one side, (for back-thinning to a special surface-lacquer protected), or both sides by inverting the sample after jet polishing about half way thru it. Microshield lacquer, (from South Bay), works great to protect surfaces from etching, dissolves in acetone, and may be thinned to reduce shrinkage when thinning soft, annealled copper for example. Also, the entire 3 m.m. disc surface can be polished via a 3 m.m. jet; using an external "timer/switch",and a D.C. power supply, planar "sectioning" of as little as 100 nm. can be removed from a surface. The jet polishing electrolyte and conditions will work. The large jet can be used to etch a surface for optical photos by simply reducing the "polishing" voltage about 20% for a couple of seconds! The 300 volt, 150 mA. capacity power supply exeeds other manufacturer's units and makes use of non-acid "BK-2" type electrolytes possible-a must for many materials. The line-of-sight optical shut off system can be fitted with a variety of color spectrum light sources for special uses. The standard infra red LED and detector bias may be independently adjusted to give the desired sensitivity setting. It will make electron transparant regions in pure annealled metal such as aluminum-with no hole! Of course the setting is normally set for a 20 micron hole with a very thin edge (quite reproducible, of course). Alignment of the parts is easy and stays set a long time. Even saphire light pipes are available for hydrofluoric acid or bromine/alcohol solutions. PVC plastic parts are available and may be substituted for metal ones for such strong chemical baths. Low temperatures of -50 degrees C. are no problem. The sample is accesible for rapid rinsig after swinging the detent-equipped jet support to one side. In 25 years of use, these instruments have saved one man per year in labor cost, (roughly $100,000/yr.), or $2,500,000--due to the ease and speed with which excellent samples can be made. About 90 to 95% of the samples attempted are good once- conditions are established. In my opinion, all the jet polishers have improved with time, but the South Bay 550 C and 550 D units are unmatched when it comes to working with the newer, difficult materials which should be viewd DURING thinning. They permit me to thin about 300 TEM foils/year in my spare time.
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Does anyone know if we'll be having, and if we should bring, examples of printer output from various printers at the meeting this year? I think it has been an excellent method for quickly comparing the various printers, and since they are ever upgrading the technology and we go right along buying new printers it seems like a very reasonable thing to bring some examples along and lay them out in the computer room again, eh?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Nina Stromgren Allen Professor, Department of Botany Box 7612 North Carolina State University Raleigh, NC 27695-7612 Phone: 919-515-8382 (Office), 515-3525 (Lab), 515-2727 (Department Secretary) Fax: 919-515-3436
I see a potential problem brewing here and the hint at tempers going up. It is time to end this particuliar thread in the public forum. This is not the place to carry out long winded commerically related arguments. If you have a problem with this send a private message to me.
COMPUTER INTERACTIVE HIGH-RESOLUTION TRANSMISSION ELECTRON MICROSCOPY
N.C.E.M, LBNL, Berkeley, California
The National Center for Electron Microscopy announces its fourth ANNUAL SUMMER SCHOOL on COMPUTER INTERACTIVE HIGH-RESOLUTION TRANSMISSION ELECTRON MICROSCOPY, including Image Acquisition, Image Processing and Image Simulation, to be held at the National Center for Electron Microscopy during the week of August 25-29, 1997.
The aim of the School is to train participants in the techniques of computer-assisted high-resolution electron microscope image acquisition and image interpretation, including remote-control microscopy. Participants will learn general principles and apply them to specific cases. Participants will be taught the use of computers to obtain images on NCEM microscopes, followed by training in the use of application programs for image interpretation by image processing and image simulation. Participants wanting to apply school techniques to their own projects will be encouraged to extend their visit to NCEM into the next week -- note that this requires a proposal be submitted with advance notice sufficient for project approval.
For more information, please see - http://ncem.lbl.gov/NCEM/workshops.html
:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:. Michael A. O'Keefe, Deputy Head National Center for Electron Microscopy Lawrence Berkeley National Laboratory University of California Berkeley, California 94720 tel: (510) 486-4610 fax: (510) 486-5888 email: maok-at-lbl.gov http://ncem.lbl.gov/ :.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.
The July 1997 Microscopy Listserver Archives are now available on line at the MSA WWW Site (http://www.msa.microscopy.com). A healthy month with a file size of 1.3 Mbytes . That according to my records is the most ever posted. In case your curious a total of 1.1 Million Email messages were sent out to the members listserver from this server during the month of July.
Also just a reminder that the Microscopy & Microanalysis 97 meeting is only days away, August 10-14 in Cleveland Ohio. Hotel rooms are at a premium this year so expect a big crowd.
For those of you that can't join us this year, check the MSA WWW pages for updated information on what is happening. Last year we were able to organize a live Internet Video Feed from the meeting. Depending upon the hardware present we will try to organize something along those lines again.
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Hi, there, I am running a real time captureing for zoospore discahrge of a plant pathogenic fungus through Matrox Inspector, my attempt is to put a clip of this on our web site, the problem I am having now is the file size usually too big. Does anyone know anyway we can resize or edit the avi files into a smaller size? Please drop me your input directly into my mail address. Thanks ahead. KerChung
Can anybody out there supply me with a/the origanal reference on the use of OsO4 as a vapour fixative as for delicate biological material?
ThankyouAlan N Hall Unit for Electron Microscopy Faculty of Biological & Agricultural Sciences University of Pretoria Pretoria 0002 Republic of South Africa Tel: +27-12-420 3297 Fax: +27-12-420 3266
Re: Staining Epon with PAS & IKI. I recently got some ambiguous results wit PAS. We were staining thick (2u), epon-embedded plant sections that had been fixed in aldehyde followed by Osmium, and embedding. We knew that the tissue was lipid rich based on prior work with fresh material and staining of Osmium-treated material with Sudan Black B. The lipid bodies were circular as one would expect. We knew that the tissue could have significant amounts of starch in it, so we used the PAS protocol. Many of the circular bodies which we thought were lipids also stained positively with PAS. I decided to add some aqueous IKI to unstained sections and I was surprised to get a positive reaction which clearly showed the starch grains in amyloplasts. The grains stained brown and could easily be distinguished. The IKI was a potent mix of 1 gm. I & KI in 100 ml, and aged for a year or so. This may be a trivial note (please don't tell me that however), but I thought it might be of some interest.
Question - Might the positive PAS reaction by lipid droplets be due to glycolipids or is this a false positive reaction? I have stained lipid-rich plant material before and have never seen a PAS response like this.
Larry Glitch wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi everyone! } } I heard - although I can't believe it - that RJ Lee was giving away an } SEM at the MSA meeting. Am I nuts or has anyone else heard about this? } } Thanks! } } Larry
Dear Larry;
YES! it is true RJ Lee Instruments Ltd is giving away (for a 90 day trial period) a Personal SEM at the M&M Conference this year. We are sponsoring a competition asking participants to bring a sample to the RJ Lee booths (400, 402, 404) and take a picture using the PSEM. The best photo will win. Contest rules are available on request. Anyone interested in signing up for some time on the instrument can call Doris Allison (800-573-PSEM) and make an appointment. You can also send an email message and I'll see you get on the calendar. If you are not familiar with the PSEM or Computer Controlled Electron Microscopy, please come see us.
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i'm currently using spurr's to embedd metal powders (currently Al) with thin oxide layers on the surface (i.e. Al2O3) ...the current protocol is to flat embed the powders on an aluminum weighing dish using spurrs + 1 drop Z60/40 and on the next day...beem capsule embed slices of the flat embedded samples cut with a razor blade...then proceed to microtome for SEM and TEM observation
i'm not sure if the problem is with the sample preparation...should i try a different embedding media?? or if it is with the actual microtoming procedures..
i've managed to get a few good sections but would like more consistency and improvement on the quality of the section...any suggestions would be greatly appreciated....
many thanks in advance
Sincerely, Michael Mandanas Particulate Materials Center Penn State University University Park, PA USA mxm67-at-email.psu.edu
As you can see by now a JUNK Email/Marketing program has discovered the Email address of the Listserver. I have contacted the organization that is running this "service". Please ignore the message that concerns "removal request" you are NOT being removed from the Micrsocopy Listserver. Rather supposedly the Microscopy Listserver is being removed from their system. They unfortunately have a system which sends out the notification to each Email address.
I can't do anything further about it at this time. Let's see how well their "system" works.
We are shopping for a TV rate camera to interface to our Hitachi H-600 TEM via a 35 mm camera port (scope is equipped with STEM). We plan to use this for teaching and when more than one person (i.e. operator and researchers) are working at the scope, not for acquisition of high-quality digital images. To the best of my knowledge these systems work as follows: a small phosphor screen will be moved into the beam path, a camera is focused on this screen and the image is displayed on a small B&W monitor. I have found two vendors so far (Fullam and Gatan), but our purchasing department wants more. If you know of any other vendors for this kind of equipment - or are vendors of it - please reply directly to me by e-mail.
Thanks in advance for any replies.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Please post the following job description for our company on your employment page. Thank you for your help.
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TECHNICAL SUPPORT AND SALES REPRESENTATIVE
Supplier of high quality instruments and digital imaging products to the domestic & international microscope market, is looking for a motivated person to provide technical & sales support. A BS in biology, medical technology or other sciences is required. We are introducing new products and opportunities exist in this rapidly growing company. Excellent benefits include choice of fully paid PPO or HMO medical plans, dental, prescription, vision, life ins., 401K and educational reimbursement.
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Best Regards, Bill Solinski
Diagnostic Instruments 6540 Burroughs Sterling Heights, MI 48314 Phone: 810-731-6000 FAX: 810-731-6469 Website: www.diaginc.com e-mail: info-at-diaginc.com
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Yes we are putting our images on CD ROMs in various Photoshop formats and reading them on both Power Mac/Macs and PCs - no problem. We haven't gone to the multiple session, do it yourself CDs but we have someone here make them on a higher quality, dedicated system, presumably because it is more reliable in giving us an error free disk. Also I disagree about Zip drives, we use them to transfer large numbers of images from our lab to our customers; however, I've been told by computer sales that Zip drives are discontinued. Any confirmation? If so, that goes along with a previous message about archiving digital images when the hardware won't be around to read the data. I have already run into this with data stored on 8 1/2" floppies for a Kevex EDXS instrument that is no longer around. Here is a business opportunity for someone who wants to archive all this old equipment to read archived data; of course, you'll need extra units for spare parts....:-)
Hope this answers your question, Tom.
Damian Neuberger neuberd-at-baxter.com
Our multi-user facility is currently archiving our confocal and LM digital images on Panasonic optical disks (re-writable, very stable -at- about $125 for 1 GB). The disadvantage is that few of our users have their own Panasonic drives so most people simply archive the images at our core and then move the ones they want by FTP as needed. I would like to switch to a more universal medium - namely CD ROM's. My understanding is that CD's can now be written to in multiple sessions so you don't need to fill an entire disk at once. Furthermore, it is my understanding that a disk of TIFF images should be readable by both IBM/WINTEL and Mac/PowerPC types computers. Is anybody actually doing this? Comments on how reliable are the recorders, which ones are best, pitfalls, etc would be appreciated. Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that route since that they are not as ubiquitous as CD drives. Thanks in advance.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Michael P. Mandanas and microscopists: You are not describing the actual problem, but I expect that the Al particles pull out of the sections. Use the hardest mixture of Spurr's and over-cure a bit. Use a small block-face and a diamond knife, best one with a more obtuse angle than the biology knives have. For SEM specimens could be more effectively ground. Do not use carborundum powder because particles will be embedded within your specimen. Diamond paste would be a lesser problem but best are diamond lapping films (from EMS or ProSciTech), which are plastic films with embedded, not glued diamond particles. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} i'm currently using spurr's to embedd metal powders (currently Al) with } thin oxide layers on the surface (i.e. Al2O3) ...the current protocol is to } flat embed the powders on an aluminum weighing dish using spurrs + 1 drop } Z60/40 and on the next day...beem capsule embed slices of the flat embedded } samples cut with a razor blade...then proceed to microtome for SEM and TEM } observation } } i'm not sure if the problem is with the sample preparation...should i try a } different embedding media?? or if it is with the actual microtoming } procedures.. } } i've managed to get a few good sections but would like more consistency and } improvement on the quality of the section...any suggestions would be } greatly appreciated.... } } many thanks in advance } } Sincerely, } Michael Mandanas } Particulate Materials Center } Penn State University } University Park, PA USA } mxm67-at-email.psu.edu } }
I am working with decalcified bone which has been fixed with osmium and embedded in Spurrs for TEM. There are sections being taken for LM which we would like to do optical staining on but I am encountering alot of difficulty.This is not my field of study, I am a work-study student and would appreciate any help/suggestions for the following:
Semi-thin and ultra thin section of decalcified bone (14% EDTA) which is fixed in osmium and embedded in Spurrs. I have tried a 2%NaOH/absolute alcohol to deplasticize then H2O2 to remove the osmium and have been working with Villnueva trichrome bone stain and tetratchrome bone stain. The specimens are only picking up a very small amount of stain after 24 hrs of staining and they are degraded (probably from the H2O2).
If anyone has further suggestions please let me know. Thanks.
i apologize for forgeting to state the actual problem...i guess things done in haste does go to waste. anyway, what i forgot to mention in my previous email is that the particles seem to pullout and the spurrs does not hold the particles strong enough during sectioning-there are void spaces around the particles and the surface is rough...another problem we've encountered is charging on the TEM when attempting to get higher magnification images - a spurrs stability under electron beam problem? i hope this clarifies my problem and again, i apologize for the confusion
sincerely
Michael Mandanas Particulate Materials Center Penn State University University Park, PA USA mxm67-at-email.psu.edu
On July 31 Ian Laren asked: } Also, does anyone have any idea why oily blobs (obviously from oil in the } vacuum system) sometimes appear on the sample in the very place that you } are observing (as opposed to any other place on the sample)? } } Thanks }
Ian and all:
The probable mechanism is that oil vapor molecules are ionized by the electron beam (Mass spectrometers use this ionization method), and then the positive ions are attracted to the negative charge deposited by the beam on the sample. Thus a hydrocarbon polymer is deposited exactly on your area of interest.
A long discussion of contamination and its control appears at our web site.
Ronald Vane XEI Scientific SEM-CLEAN anti-contamination systems (650) 369-0133 http://www.msa.microscopy.com/SM/XEI/XEIHomePage.html
Recently, I attempted to obtain 70 nm sections from a specimen embedded in Spurr's. The plastic was separating from the tissue shortly after the cut. Furthermore, these sections are not holding up to the beam. I believe my problem is an incomplete infiltration. I allowed the blocks to sit in a 90 degrees C oven over the weekend to see if that would help. The problem, however, continues to exist. Is anyone familiar with any methods of depolymerization and reinfiltration of embedded specimens? Any other suggestions that may help would be greatly appreciated.
I prferred Epon to Spurrs because the hardness of the Epon can be adjusted readily by different ratio's of mixtures A and B as described in the Biology handbooks. Look into any biology related microscopy book to get the proper ratios. I think you will find that the epon is much harder. I don't know if that will solve your problem. As for the charging, try a thin coating of carbon before inserting into the TEM.
Good luck.
Roberto R. Garcia EMF Manager Wright State University
Michael Mandanas wrote: ======================================= i'm currently using spurr's to embedd metal powders (currently Al) with thin oxide layers on the surface (i.e. Al2O3) ...the current protocol is to flat embed the powders on an aluminum weighing dish using spurrs + 1 drop Z60/40 and on the next day...beem capsule embed slices of the flat embedded samples cut with a razor blade...then proceed to microtome for SEM and TEM observation
i'm not sure if the problem is with the sample preparation...should i try a different embedding media?? or if it is with the actual microtoming procedures.. ================================================= Our laboratory has been diamond knife thin sectioning metal powders, including those of aluminum for over twenty seven years. I would like to summarize our experiences, some of which actually led to some new product concepts (e.g. the concept of a materials science diamond knife):
1] Aluminum is a special case because relative to most other powers one might want to section, it is very soft. If it further "special" because if there is indeed an oxide layer, e.g. like a layer of anodization, since the adhesion of the oxide layer is not the greatest, one has to use vacuum embedding. Otherwise, the "pores" of the oxide layer do not get impregnated with the resin and there is then an extraordinary level of "pullout" of the particles.
2] After trying all of the various embedding resins, we have settled on our own SPI-Pon(TM) resin kit. We suspect, but don't know it for a fact, that at least some of the so-called "Epon(TM) 812 replacement" kits offered by others would work just as well. It seems like one has not only the maximum ease with which the hardness can be controlled, but for the hardest of powders, it seems, at least to us, that this resin system permits the curing of the hardest possible block, yet when sectioning, there is less of a tendency to "chatter" and "compress".
3] Diamond knives must be used in this kind of application, and trying to use glass will just turn out to be a grand exercise in frustration. And further on that, we would certainly want to be using a "materials science" diamond knife. Now at the risk of sounding "commercial" I would like to say that there are three schools of thought on the matter of "materials science" diamond knives: a) The concept itself is a gimmick, b) make your knife with an angle that is much more blunt (e.g. not such a sharp angle), or c) finish off the knife with the same angle as is used for life science knives generally (e.g. 45 deg.), but perhaps not worry about the last of the fine striations, since any that are there are going to be small indeed compared to the larger ones that will be put in on the first slice.
We ourselves are part of the last school of thought on this issue. It is certainly not a gimmick because why would anyone in their right mind pay top dollar for the perfect edge, only to reduce it to a state that is no better, and probably worse than a "materials science" knife from that third school when it is brand new? The concept of using a more blunt of an angle on the knife, while in theory (and usually in practice) such an edge will last longer, because of the less "sharp" edge, compression effects tend to be greater as well as also, the instances of particle "pull out". Sometimes these effects are sufficiently profound that usable sections can not be made at all (it depends on the powder being sectioned). And in the end, if usable sections are obtained, usually more sections have to be made, putting more wear and tare on the knife edge, so that in the end, its useful lifetime is not all that much longer.
4] We have found the above to be true, not just with the relatively spherical metal powders, but also with aluminum flake, of the type used in automotive (metallic) paint coatings.
While there is no question there is some "art" (as well as experience) involved here, once the "secret" is known, it is no longer magic. On the other hand, I have suspicions that there are some who have obtained quite excellent sections using other embedding resins and perhaps diamond knives from the second school as well, so even we won't claim to know all of the "magic". I can, however, speak only from our own experiences.
Disclaimer: Our firm offers diamond knife thin sectioning,on these kinds of materials, as a service, and for a fee for others and we also offer the SPI Materials Science line of diamond knives for persons wanting to do this type of work themselves. Information about these products and services can be found at the website address given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
At 08:27 PM 8/4/97 -0400, Bill wrote: } I have downloaded your database files but cannot unzip xray_ms.zip, it gets } a crc error when I try. I downloaded xray.xls and xray_ms.xls but neither } would open with Excel 7.0 on a PC. Can you offer any suggestions? } I'm getting similar feedback from others ... and I've just tried to open the files without luck (... weird ...). I've just opened the working spreadsheet (Excel v.7). It actually is 2 sheets ... one of which is the database and the other sine-theta values for Cameca spectrometers, which can be easily be converted if you change the cystal 2D and k values, and the sine-theta range. It could also be easily modified for LiF and wavelength geared spectrometers (Note: the sine-theta sheet has hidden columns and frozen rows for visibility). I will zip it then unzip it for a check, and let you and others know I can dump the separate sheets as other types of files or text files. The zipped Excel (v.7) spreadsheet xrayxls7.xls (xray_ms.zip) will be available via anonymous FTP at whitewater.uoregon.edu/share/cameca/ ... the other types of files can be requested.
cheerios, shAf
BTW ... the 2Mb zipped file tested, unzipped and loaded A-OK ... the XLS file also exists at the FTP site but it is 6Mb. Let me know if there are any problems ... I've also been successful in creating a PDF file but I'm waiting to get the 2D and k values for one more xtal before it is finished.
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
The Iowa Microscopy Society meeting is to be held on September 25th and 26th, 1997. An immunocytochemistry workshop is being offered, and advance registration is suggested. More information on the workshop and the Iowa Microscopy Society's meeting can be found at http://www.uiowa.edu/~cemrf/cemrf/ims_announce.html or you can give Kenneth Moore or myself a call at 319-335-8142.
Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
recently i have been studying polymer distribution with ceramic powders (in this case Al2O3)...in one case, i stained polyvinyl alcohol with RuO4, and there is a gradient of the polymer distribution in the powder...now my question is, what would be the best way to determine and/or calculate the concentration gradient / distribution of the polyvinyl alcohol...does the RuO4 concentration directly translate to polymer concentration? ..if so, can i just take sections and do mass spec each of the sections?? should i be working with a different staining agent for PVA, what about other polymers like polyethylene glycol, acrylics ?? a more general question would be, what is a recommended refernce for polymer staining..i've read Linda Sawyer's "Polymer Microscopy" textbook but i was looking for a more detailed discussion...if mechanisms are included, the more help it would be... any suggestions would be greatly appreciated...
thanks again in advance sincerely
Michael Mandanas Particulate Materials Center Pennsylvania State University University Park, PA 16801 USA mxm67-at-email.psu.edu
Updates for both Mac and Windows versions are now ready for downloading at http://Members.AOL.com/ImagProcTK/update.htm
Intensity Profile is now available for the PC, Autocontrast for both platforms expands contrast over the full range of the image (great for images with shading, or TEM sections with varying thickness), and there is a spreadsheet with examples for data analysis (statistics, graphs, and sphere unfolding).
DIRECTIONS FROM CLEVELANE HOPKINS INTERNATIONAL AIRPORT TO DOWNTOWN CLEVELAND VIA THE RAPID TRANSIT AUTHORITY (RTP-RAPID)
From the baggage claim level at the airport, proceed down one level via the escalators in the center of the baggage claim. Continue following signs to the rapid transit station. Exact change is required - $1.50. Take the rapid transit to the Tower City - Downtown Terminal which is the last stop. Proceed through the turn styles.
If your accommodations are at the Ritz Carlton, once through the turn styles, turn right and proceed up the two sets of short escalators. The entrance to the Ritz Carlton is on your left.
If your accommodations are at the Renaissance Cleveland Hotel, once through the turn styles, proceed left up to the long set of escalators. At the top, make an immediate left and then another immediate right and follow the signs towards Public Square. Once you reach the Disney store, turn left and continue straight through the double set of glass doors. Proceed up the staircase and you are in the lobby of the Renaissance Hotel.
If you are staying in another downtown property, follow the directions as listed above for the Renaissance Hotel however, at the Disney store, proceed straight through the glass doors heading outside where you will find taxi's.
The Marriott and Sheraton are 2-1/2 and 4 blocks respectfully if you choose to walk. Once outside walk across Public Square towards the Key Bank Center where the Marriott is located. The Sheraton is within sight from the Marriott entrance, next to the Cleveland Convention Center.
{P} {B} DIRECTIONS FROM CLEVELANE HOPKINS INTERNATIONAL AIRPORT {/B} {BR} {B} TO DOWNTOWN CLEVELAND VIA THE RAPID TRANSIT AUTHORITY {/B} {BR} {B} &n bsp; (RTP-RAPID) {/B}
{P} From the baggage claim level at the airport, proceed down one level via the escalators in the {BR} center of the baggage claim. Continue following signs to the rapid transit station. Exact change is {BR} required - $1.50. Take the rapid transit to the Tower City - Downtown Terminal which is the {BR} last stop. Proceed through the turn styles.
{P} If your accommodations are at the Ritz Carlton, once through the turn styles, turn right and {BR} proceed up the two sets of short escalators. The entrance to the Ritz Carlton is on your left.
{P} If your accommodations are at the Renaissance Cleveland Hotel, once through the turn styles, {BR} proceed left up to the long set of escalators. At the top, make an immediate left and then another {BR} immediate right and follow the signs towards Public Square. Once you reach the Disney store, {BR} turn left and continue straight through the double set of glass doors. Proceed up the staircase and {BR} you are in the lobby of the Renaissance Hotel.
{P} If you are staying in another downtown property, follow the directions as listed above for the {BR} Renaissance Hotel however, at the Disney store, proceed straight through the glass doors heading {BR} outside where you will find taxi's.
{P} The Marriott and Sheraton are 2-1/2 and 4 blocks respectfully if you choose to walk. Once outside {BR} walk across Public Square towards the Key Bank Center where the Marriott is located. The {BR} Sheraton is within sight from the Marriott entrance, next to the Cleveland Convention Center.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello all, } } Recently, I attempted to obtain 70 nm sections from a specimen embedded in } Spurr's. The plastic was separating from the tissue shortly after the cut. } Furthermore, these sections are not holding up to the beam. I believe my } problem is an incomplete infiltration. I allowed the blocks to sit in a 90 } degrees C oven over the weekend to see if that would help. The problem, } however, continues to exist. Is anyone familiar with any methods of } depolymerization and reinfiltration of embedded specimens? Any other } suggestions that may help would be greatly appreciated. } } Sincerely, } } Dan Caruso c/o Eugene Gordon } } Your best bet would be to start over. However I was faced with a similar problem two years ago with some tissue which had been very poorly infiltrated (in another lab)and brought to us for evaluation. You need patience: 1. Cut the tissue out of the existing blocks. Trim as closely as possible. 2. Expose the tissue to numerous changes of propylene oxide in vials which are on a rotator. You will see the unpolymerized resin gradually enter the PO. Keep changing fluids. This may take 48 hours. Tissue will get softened. When you feel that no more changes are forthcoming, reembed the tissue in the same protocol as the first, making sure that each time the tissues remain in motion. Lengthen the infiltrations steps, especially the first one which is likely to be mixture of resin and intermediate agent. Reembed and polymerize and hope. The original polymerization may have gone far enough so that the above method is not successful. The whole procedure may take a week, but if the redoing is successful you may save valuable tissue. If the method is not adequate for sectioning, you must start over. If the sections are "delicate" pick them up on formvar coated grids and carbon coat them. 3. What was the cause of this problem? It may have been water! Was propylene oxide or acetone, or alcohol left in the tissue? Or was it truly only inadequate infiltration??? It is important to make a distinction. Good luck. Hildy
Email: hyphae-at-msn.com Name: Scott Mcphee School: Santa Rosa Junior College, California
Greetings,
I am a beginning Mycology and Phytopathology student and am looking for a good quality inexpensive ( {$400) microscope. From the prices I have seen while browsing various sites it looks like this will probably be used due my limited funds.
I would like to be able to spore and cellular characteristics while I am at home. I can take a couple items to school and look at them there, but I often have large amounts of things to study and it would be much more conveniant to be able to do this at home. The fungi often turn to mush also before I can get them to the lab at school.
Could you reccomend a power range and perhaps a model?
I was given a block of paraffin wax with specimen embedded in it. I would like to know how can I get away the wax to get out the specimen. It is rather urgent and I would appreciate immediate reply if possible. Thank you very much.
Many people have problems obtaining information easily from their TEM and=
SEM, I see it almost every day. The area of instrument set up that causes=
more of these problems than any other is that of filament position.
Most people set the filament a long way from the cathode aperture and thi= s leads to a long filament life but a low emission current. Moving the filament forward increases emission current but decreases filament life. =
With care and the additional use of the bias or emission control this forward movement of the filament will result in an improvement in performance - resolution. One problem, people place more credence in having a long filament life than in getting more from their microscope! =
Many buy a new machine because they feel the reason they cannot obtain th= e result they want is down to the microscope. Forget filament life if you want more from your machine, in my experience an instrument will be transformed if you push the gun a little harder. I could tell you so man= y stories where we have done this, much to the amazement of the owner and delight of the dissatisfied customer!
As for changing filaments the more you do it the less frightening it becomes, its really not difficult and to be honest if you want more from your microscope the cost is very little.
Why should people calibrate their microscope you may ask? If you do not take what is a pretty simple step how do you know that the instrument is working correctly? Is it not better to test the microscope via resolutio= n, contamination and calibration than to have a hoard of customers banging o= n your door complaining that their results are poor; too late! Preventativ= e maintenance, spotting problems at higher levels of operation than the normal in your laboratory, should be part of every well run laboratories routine. Spot the problems before they become a disaster and get them fixed before anyone else notices, thats they way to run a unit! It is n= o good saying the engineer does it one or twice a year, (does he really?) that gives a long period of uncertain operation and possible problems. I= n addition, if you learn to operate your microscope at higher levels than i= s the norm you improve your own techniques and powers of observation.
On one of my hobby horses, I believe that every laboratory should test it= s instruments and its operators routinely to see how they all perform. If you set a standard where you are testing in this way you instantly raise the levels of expertise and have standards which may be used over many months to prove that the laboratory is moving forward. Without such a regime laboratories stagnate! Everyone feels they do a great job but wit= h no form of standard how do they know? We talk about Quality in electron microscopy in relation to how we set analytical standards and calibration= ; good! But an even more important question which is totally ignored is "how good are the operators?". I have said before microscopists are supposed to be scientists, real scientists would constantly test themselves!
Many people have problems obtaining information easily from their TEM and=
SEM, I see it almost every day. The area of instrument set up that causes=
more of these problems than any other is that of filament position.
Most people set the filament a long way from the cathode aperture and thi= s leads to a long filament life but a low emission current. Moving the filament forward increases emission current but decreases filament life. =
With care and the additional use of the bias or emission control this forward movement of the filament will result in an improvement in performance - resolution. One problem, people place more credence in having a long filament life than in getting more from their microscope! =
Many buy a new machine because they feel the reason they cannot obtain th= e result they want is down to the microscope. Forget filament life if you want more from your machine, in my experience an instrument will be transformed if you push the gun a little harder. I could tell you so man= y stories where we have done this, much to the amazement of the owner and delight of the dissatisfied customer!
As for changing filaments the more you do it the less frightening it becomes, its really not difficult and to be honest if you want more from your microscope the cost is very little.
Why should people calibrate their microscope you may ask? If you do not take what is a pretty simple step how do you know that the instrument is working correctly? Is it not better to test the microscope via resolutio= n, contamination and calibration than to have a hoard of customers banging o= n your door complaining that their results are poor; too late! Preventativ= e maintenance, spotting problems at higher levels of operation than the normal in your laboratory, should be part of every well run laboratories routine. Spot the problems before they become a disaster and get them fixed before anyone else notices, thats they way to run a unit! It is n= o good saying the engineer does it one or twice a year, (does he really?) that gives a long period of uncertain operation and possible problems. I= n addition, if you learn to operate your microscope at higher levels than i= s the norm you improve your own techniques and powers of observation.
On one of my hobby horses, I believe that every laboratory should test it= s instruments and its operators routinely to see how they all perform. If you set a standard where you are testing in this way you instantly raise the levels of expertise and have standards which may be used over many months to prove that the laboratory is moving forward. Without such a regime laboratories stagnate! Everyone feels they do a great job but wit= h no form of standard how do they know? We talk about Quality in electron microscopy in relation to how we set analytical standards and calibration= ; good! But an even more important question which is totally ignored is "how good are the operators?". I have said before microscopists are supposed to be scientists, real scientists would constantly test themselves!
} I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much.
Would your specimen be harmed if you used solvent to remove the paraffin? Hexane, heptane or Petroleum Spirits 60-80, warmed slightly if necessary, would be suitable. They are not very toxic (but don't breathe too much), but they are highly flammable.
Several washings in a small quantity of solvent each time are MUCH MORE EFFICIENT than washing in one big lot of solvent (Bunsen's dilution law).
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
IF the speciment can take xylene, and wasn't directly embedded in paraffin, then:
Cut the specimen out of the wax, trim away as much wax as possible, then soak in xylene or a xylene replacement (like HistoClear). Make several changes to make sure you get all of the wax out. Times depend on specimen size and nature. 3 X 10 minutes might work, or might be too short. Look for a transparent quality to the specimen, that should indicate that all the wax is gone and completely replaced with xylene.
What you do next depends on what you're after. The specimen can be backed down through a xylene-alcohol series to 100% alcohol, and then through alcolhols to 70% EtOH, or even to water. Just reverse the usual schedules using in LM paraffin staining.
Whether the specimen can go through this and stay in good condition depends on the specimen, and to some extent on how much you need fine details. Most can, but not all (I don't have any examples in the top of my head).
However, before you de-embed, what do you what to look at? If the specimen is say, a whole insect, you can trim away the excess wax, and then, while still embedded, "dissect" the specimen by carving away the unwanted bits. The wax holds the specimen together, and you can carve on the curve, instead of just flat planes like a microtome does. (Also, the microscope lights [use fiber optics] melt the wax locally, and the streams of molten wax wash away the debris.) After carving, then proceed through the series as above.
Phil } } } I was given a block of paraffin wax with specimen embedded in it. I } } would like to know how can I get away the wax to get out the specimen. } } It is rather urgent and I would appreciate immediate reply if possible. } } Thank you very much. } } } } } } } } Catherine Tang }
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{(((( Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again *****
What do you want to do with the specimen? Two quick ways to remove paraffin- 1) Melt it away in a 65 degree oven and 2) Dissolve it away with xylene. More info is needed to answer your question.
Ed Calomeni Dept Pathology Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
We deparaffinize with xylene. The time required will depend on the size of the tissue, so I suggest you cut out a small piece if possible. We then rehydrate step wise to water. Then we handle it as a normal TEM specimen. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 02:10 PM 8/6/97 +0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
The most straight forward way to remove the paraffin would be to trim the block as close to the tissue as possible and place it in xylene or histoclear to dissolve the rest of the paraffin away. I would do at least three changes, the time of the change would depend on the block size, then rinse out the paraffin/xylene with 3 changes of 100% ethanol and process in your new media, starting from the 100% ethanol step. I have done this with some success, though often the tissue that's processed for LM doesn't look so hot at EM. That depends on how it was fixed.
Good luck,
Karen Pawlowski PhD student/Histology Tech. UT Dallas/ UT Southwestern Medical Center Dallas
On Wed, 6 Aug 1997, Tang Ee Koon wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much. } } } } Catherine Tang }
Hello everyone, I am looking for a list of vendors of used light microscopes and parts. I would like to have their phone number or other information that will help me contact them. You may reply directly to me. Thank you Sincerely Karpura kkommine-at-mbl.edu Marine Biological Laboratory Woods Hole
We have been using calibration tolerances of + or - 5% for both our magnification and spectrometer calibrations for quite some time now. Apparently these values have been established through supplier and user inputs quite some time ago. I am curious as to whether these values are consistent with those currently used or whether they are too loose. We are ISO 9002 registered and quality measures are very much a concern. TIA
} I am a beginning Mycology and Phytopathology student and am looking for a } good quality inexpensive ( {$400) microscope. From the prices I have seen } while browsing various sites it looks like this will probably be used due } my limited funds. } } I would like to be able to spore and cellular characteristics while I am at } home. I can take a couple items to school and look at them there, but I } often have large amounts of things to study and it would be much more } conveniant to be able to do this at home. The fungi often turn to mush also } before I can get them to the lab at school. } } Could you reccomend a power range and perhaps a model?
You'll be able to get by with 10 & 40x objectives, but 100x (oil immersion) would be nice, and maybe 4x for the big stuff. 10x eyepiece. A condenser is highly desirable. Try the "surplus" departments at U.C.S.F. and U.C. Berkeley for some really good buys. Take a friend who knows scopes with you if possible. Ask your college to buy the U. of Washington CD-ROM listed in the MICRO bibliography (address below).
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Tang wrote regarding retrieval of specimen from wax} Simply cut down the wax to the size of the specimen and place in several changes of xylene (could be warmed slightly in a hood). Assuming you wish to attempt TEM microscopy on the specimen at this point once all the wax is removed you don't need to dehydrtate since the specimen is already dehydrated and attempts to use osmium tetroxide seem futile. Next process in propylene oxide and infiltrate with plastic as usual and stain the heck out of the grids. Results are usually very poor to marginal but diagnoses have been made on some tumors this way.
fhayes-at-dow.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Does anyone know of a vendor that supplies lacey carbon coated Be grids? } } If not, are there any suggestions out there regarding how to make such a } substrate? } } Fred Hayes } The Dow Chemical Co } Analytical Sciences Laboratory } 1897 Bldg., E78 } Midland, MI 48667 } 517-638-2203 } 517-638-6443 fax }
Dear Fred, Ladd Research has done lacey carbon coated Be grids in the past for customers. Please contact us via e-mail or 1-800-451-3406 and we can discuss pricing and what size mesh you would like it done on.
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Clean UA/lead stain 8/6/97 12:49 PM
Has anyone heard of using a few drops triton-X detergent in aqueous uranyl acetate to help keep stain clean when staining with uranyl acetate and lead? If so, does anyone have a logical explanation why this may work? Thanks in advance for the help. Linda Chicoine Center for Cell Imaging Yale University New Haven, CT
I've been ignoring the microscopy meeting that is coming to Cleveland and suddenly my boss said to me today, "hey, we should think about going to this thing next week". We work in Cleveland so thats not a problem. Could someone send me information like schedules and fees and seminars and classes, etc, etc, etc. Thanks. Mark Darus
Intel Corporation currently has an open position for a SIMS technician at its Santa Clara site's Materials Technology department. The SIMS group supports the Santa Clara site's development and fabrication facilities that produces state of the art microprocessors. The Materials Technology department's scope encompasses the whole process from silicon to a packaged device that includes process trouble-shooting, transfer and equipment qualifications, device failure analysis/fault isolation and new materials development to name a few. The department has a wide variety of state of the art instrumentation, including: SIMS, Auger, TEM, SEM/EDS, FIB, AFM, XRF, ICP-MS, FTIR, Raman, Pico and Nano Indentation.
Job Description:
The technician opening involves second shift operation of Quadrupole (Atomica) and Magnetic Sector (Cameca IMS- 6F) SIMS tools. SIMS is an Ultra High Vacuum analytical tool which uses ion beam sputtering combined with mass spectrometry detection to analyze dopant and contamination levels and distributions in patterned and unpatterned wafers. Duties will include data collection, data processing, report writing, interfacing with other engineers and customers, communicating results, and workflow duties to keep the lab running smoothly. The successful applicant must be self-motivated and capable of working with minimal supervision.
Qualification: An AA or BS degree with Engineering or physical science background or equivalent experience is required. Ability to work in a team oriented environment and good communication skills are critical. Experience with analytical equipment or Ultra High Vacuum or Vacuum systems is desired. Experience with SIMS and knowledge of Semiconductor processing methods is a definite plus.
Intel's industry leading total compensation package includes a competitive salary, stock options, annual employee bonus plan, bi-annual employee cash bonus payout, periodic paid sabbatical leave, and retirement plan. Intel is an equal opportunity employer.
Send resumes to or for more information contact:
Gabi Neubauer or Jerry Hunter Intel Corporation 2200 Mission College Blvd., M/S: SC2-24 Santa Clara, CA 95052
Phone: (408) 765-2241 or 765-2316 FAX: (408) 765-2393
Peter Tarquinio Evex Analytical 857 State road Princeton, NJ 08540
----------
I've been ignoring the microscopy meeting that is coming to Cleveland and suddenly my boss said to me today, "hey, we should think about going to this thing next week". We work in Cleveland so thats not a problem. Could someone send me information like schedules and fees and seminars and classes, etc, etc, etc. Thanks. Mark Darus
One of the major problems with crystals and crystal lattice specimens as = a TEM test specimen is the need to determine the level of astigmatism in th= e image. As a novice service engineer needing a crystal lattice resolution=
picture you soon learn to place a little astigmatism in the image and hop= e! Its much easier than trying to find why you cannot resolve the lattice.
The best TEM test specimen to cover a wide range of operator levels is th= e holey carbon film. Tom Mulvey stated that "the minimum discernable fring= e level is similar to the point to point resolution of the instrument"
A holey carbon film at 200,000X is a test for anyone, the object being to=
obtain the finest fringe that is even all round the hole. It tests the operators skills and their ability to truly observe an image. A through focal series is required but it is a waste of time doing this within two hours of switching on the kV as this will take time to stabilse. I have discussed heat gained in the tank needing to equal heat lost in order to obtain stability within the past few months on e-mail.
Correct the astigmatism at double the photo mag and then at the photo mag=
set the focus so that you just see an over or under focus fringe. Take a set of pictures through focus and then measure the centre of the black fringe to the centre of the white fringe ON THE NEGATIVE but only if the fringe is even. Fringes tell you a good deal about the instrument and its=
alignment (Reference Monitoring & Maintaining the TEM) Most people start=
off with about a 1 to 1.5nm resolution and if you are really good you may=
attain 0.45nm at which point the carbon structure starts to interfere wit= h the fringe.
What do you learn. =
1) You will see if the high voltage is stable - it is not perfect if you=
get focus drift, if you do not see some focus drift I would be surprised.=
2) You will notice specimen drift when you first insert the rod 3) You will realise the importance of a good anticontamination device as the hole shrinks in size. 4) You will probably need more current - its that filament position again= ! 5) How difficult it can be at this level to correct astigmatism, you are looking for a fringe like a piece of cotton not a ships hauser. 6) Typical settings - 20-25uA emission, 0.5 to 1micron spot size, CII overfocus, eucentric point, 4 second exposure to test the machine (0.5 second exposures test nothing). =
7) I do the test without an objective aperture so as to test the microscope, not the aperture cleanlyness.
Hope this helps please come back if you need more information.
Since my days as a service engineer it seems to have been accepted that o= ne could run a TEM within plus or minus 5% of the mag readout. You usually find that if the magnification is out across the range it is due to the high voltage level, specimen not at the eucentric point, or the final len= s level. If the calibration is only out after a certain point it suggest that the lens which switches in at this point is at fault.
On the SEM unless you work with perfectly flat specimens magnification accuracy is a bit of a joke! Again it seems that if you check as many instruments as I see each year that 95% are within plus or minus 10%, and=
within plus or minus 5% X to Y. Either way its usually an engineer job t= o tune the scan circuits to improve the performance.
Hello Catherine and all: Place the block in xylene and give it gentle agitation. Change the solvent a couple of times. Unless the block is very large you can de-wax the block overnight. Then dehydrate, opposite to hydration steps, but more rapidly. For the last step use single strength buffer and then fix the specimen in OsO4 and process as normal for EM specimens. Unfortunately specimens initially fixed for histology are never as good as those that were initially prepared for EM, in fact they are disappointing. However, they may show what is required and that is what matters most. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} } I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much. } } } } Catherine Tang
Hi! This is Catherine Tang. I have got a lot of replies to my question yesterday. Actually I have no experience in LM processing. I know what to do now.
Fred Hayes wrote: =============================================== Does anyone know of a vendor that supplies lacey carbon coated Be grids? =============================================== SPI Supplies has been coating Be grids for some years now. There are several "secrets", one relating to the properties and characteristics of the Be grids you want to coat. Unlike grids of Cu, Ni, and Au which are electro deposited, Be grids can be made only by etching, and different etching protocols result in grids with differing degrees of surface protruberances (e.g. those little features that tend to tear a support film). You also have to be aware that the coating of Be grids is much more time consuming than the coating of Cu, Ni, or Au grids.
The "making" of lacey carbon and followed by carbon coating is not any different than for coating grids generally.
A final inspection of representative samples of the filmed grids is mandatory before shipping off to a customer because the "yield" is so much lower (when done on Be).
You can see additional information and prices on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Some eagle-eyed environmentalists have discovered yet another Government conspiracy to conceal the true harmful nature of a chemical which commonly contaminates the environment with dire results.
Full details of this biohazard, dhihydrogen monoxide, can be found at the following website:-
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Colleagues,
Some eagle-eyed environmentalists have discovered yet another Government conspiracy to conceal the true harmful nature of a chemical which commonly contaminates the environment with dire results.
Full details of this biohazard, dhihydrogen monoxide, can be found at the following website:-
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Fred Hayes wrote: =============================================== Does anyone know of a vendor that supplies lacey carbon coated Be grids? =============================================== SPI Supplies has been coating Be grids for some years now. There are several "secrets", one relating to the properties and characteristics of the Be grids you want to coat. Unlike grids of Cu, Ni, and Au which are electro deposited, Be grids can be made only by etching, and different etching protocols result in grids with differing degrees of surface protruberances (e.g. those little features that tend to tear a support film). You also have to be aware that the coating of Be grids is much more time consuming than the coating of Cu, Ni, or Au grids.
The "making" of lacey carbon and followed by carbon coating is not any different than for coating grids generally.
A final inspection of representative samples of the filmed grids is mandatory before shipping off to a customer because the "yield" is so much lower (when done on Be).
You can see additional information and prices on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The other day I sent out directions for getting downtown from the airport via the local public transportation system, RTA. I meant to include for those who have internet browser capability the address for the RTA information system. Their site is at "http://little.nhlink.net/~rta/rtahome.html" (they also have a live picture of the Rock-N-Roll Hall of Fame). This site has all local public transportation schedules and many maps to help you get around the city using the public transportation system. The maps might also come in handy for those who are driving.
In the past year RTA has opened a new line for their rapid transit system which runs from Tower City out along the lake front to the Hall of Fame. This is their Water Front Line. I have not used this new line yet, so I don't know how to rate it.
Have a safe trip and I hope you enjoy your stay in Cleveland.
---------------------------------------------------------------- Dave Strecker mailto:dave.strecker-at-ab.com Rockwell Automation/Allen-Bradley Phone: (216)646-3250 Component Engineering ND246 Fax: (216)646-3416 1 Allen-Bradley Dr. Mayfield Hts., Ohio 44124 USA ----------------------------------------------------------------
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi! This is Catherine Tang. I have got a lot of replies to my question yesterday. Actually I have no experience in LM processing. I know what to do now.
We too are interested in calibration tolerences during our aperture QC procedures. Since the apertures we drill are as small as 5 microns and they are used in a number of applications as diverse as control jets for satelites and soldier production, tolernces can be very critical. We accept +/- 5% tolernces for magnifacation in our QC of our apertures, but I would be interested in any responses you get to this question.
We too are interested in calibration tolerences during our aperture QC procedures. Since the apertures we drill are as small as 5 microns and they are used in a number of applications as diverse as control jets for satelites and soldier production, tolernces can be very critical. We accept +/- 5% tolernces for magnifacation in our QC of our apertures, but I would be interested in any responses you get to this question.
Unsubscribe ===================================================== Marcelo Henrique Prado PEMM - COPPE/UFRJ Po.Box.:68505 Cidade Universit ria - Ilha do Fundao Rio de Janeiro-R.J. CEP.: 21941-900
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Wed, 6 Aug 1997, Tang Ee Koon wrote:
} I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much.
Would your specimen be harmed if you used solvent to remove the paraffin? Hexane, heptane or Petroleum Spirits 60-80, warmed slightly if necessary, would be suitable. They are not very toxic (but don't breathe too much), but they are highly flammable.
Several washings in a small quantity of solvent each time are MUCH MORE EFFICIENT than washing in one big lot of solvent (Bunsen's dilution law).
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Catherine and all: Place the block in xylene and give it gentle agitation. Change the solvent a couple of times. Unless the block is very large you can de-wax the block overnight. Then dehydrate, opposite to hydration steps, but more rapidly. For the last step use single strength buffer and then fix the specimen in OsO4 and process as normal for EM specimens. Unfortunately specimens initially fixed for histology are never as good as those that were initially prepared for EM, in fact they are disappointing. However, they may show what is required and that is what matters most. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} } I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much. } } } } Catherine Tang
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One of the major problems with crystals and crystal lattice specimens as a TEM test specimen is the need to determine the level of astigmatism in the image. As a novice service engineer needing a crystal lattice resolution picture you soon learn to place a little astigmatism in the image and hope! Its much easier than trying to find why you cannot resolve the lattice.
The best TEM test specimen to cover a wide range of operator levels is the holey carbon film. Tom Mulvey stated that "the minimum discernable fringe level is similar to the point to point resolution of the instrument"
A holey carbon film at 200,000X is a test for anyone, the object being to obtain the finest fringe that is even all round the hole. It tests the operators skills and their ability to truly observe an image. A through focal series is required but it is a waste of time doing this within two hours of switching on the kV as this will take time to stabilse. I have discussed heat gained in the tank needing to equal heat lost in order to obtain stability within the past few months on e-mail.
Correct the astigmatism at double the photo mag and then at the photo mag set the focus so that you just see an over or under focus fringe. Take a set of pictures through focus and then measure the centre of the black fringe to the centre of the white fringe ON THE NEGATIVE but only if the fringe is even. Fringes tell you a good deal about the instrument and its alignment (Reference Monitoring & Maintaining the TEM) Most people start off with about a 1 to 1.5nm resolution and if you are really good you may attain 0.45nm at which point the carbon structure starts to interfere with the fringe.
What do you learn.
1) You will see if the high voltage is stable - it is not perfect if you get focus drift, if you do not see some focus drift I would be surprised. 2) You will notice specimen drift when you first insert the rod 3) You will realise the importance of a good anticontamination device as the hole shrinks in size. 4) You will probably need more current - its that filament position again! 5) How difficult it can be at this level to correct astigmatism, you are looking for a fringe like a piece of cotton not a ships hauser. 6) Typical settings - 20-25uA emission, 0.5 to 1micron spot size, CII overfocus, eucentric point, 4 second exposure to test the machine (0.5 second exposures test nothing). 7) I do the test without an objective aperture so as to test the microscope, not the aperture cleanlyness.
Hope this helps please come back if you need more information.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Many people have problems obtaining information easily from their TEM and SEM, I see it almost every day. The area of instrument set up that causes more of these problems than any other is that of filament position.
Most people set the filament a long way from the cathode aperture and this leads to a long filament life but a low emission current. Moving the filament forward increases emission current but decreases filament life. With care and the additional use of the bias or emission control this forward movement of the filament will result in an improvement in performance - resolution. One problem, people place more credence in having a long filament life than in getting more from their microscope! Many buy a new machine because they feel the reason they cannot obtain the result they want is down to the microscope. Forget filament life if you want more from your machine, in my experience an instrument will be transformed if you push the gun a little harder. I could tell you so many stories where we have done this, much to the amazement of the owner and delight of the dissatisfied customer!
As for changing filaments the more you do it the less frightening it becomes, its really not difficult and to be honest if you want more from your microscope the cost is very little.
Why should people calibrate their microscope you may ask? If you do not take what is a pretty simple step how do you know that the instrument is working correctly? Is it not better to test the microscope via resolution, contamination and calibration than to have a hoard of customers banging on your door complaining that their results are poor; too late! Preventative maintenance, spotting problems at higher levels of operation than the normal in your laboratory, should be part of every well run laboratories routine. Spot the problems before they become a disaster and get them fixed before anyone else notices, thats they way to run a unit! It is no good saying the engineer does it one or twice a year, (does he really?) that gives a long period of uncertain operation and possible problems. In addition, if you learn to operate your microscope at higher levels than is the norm you improve your own techniques and powers of observation.
On one of my hobby horses, I believe that every laboratory should test its instruments and its operators routinely to see how they all perform. If you set a standard where you are testing in this way you instantly raise the levels of expertise and have standards which may be used over many months to prove that the laboratory is moving forward. Without such a regime laboratories stagnate! Everyone feels they do a great job but with no form of standard how do they know? We talk about Quality in electron microscopy in relation to how we set analytical standards and calibration; good! But an even more important question which is totally ignored is "how good are the operators?". I have said before microscopists are supposed to be scientists, real scientists would constantly test themselves!
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I was given a block of paraffin wax with specimen embedded in it. I would like to know how can I get away the wax to get out the specimen. It is rather urgent and I would appreciate immediate reply if possible. Thank you very much.
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Since my days as a service engineer it seems to have been accepted that one could run a TEM within plus or minus 5% of the mag readout. You usually find that if the magnification is out across the range it is due to the high voltage level, specimen not at the eucentric point, or the final lens level. If the calibration is only out after a certain point it suggest that the lens which switches in at this point is at fault.
On the SEM unless you work with perfectly flat specimens magnification accuracy is a bit of a joke! Again it seems that if you check as many instruments as I see each year that 95% are within plus or minus 10%, and within plus or minus 5% X to Y. Either way its usually an engineer job to tune the scan circuits to improve the performance.
Peter Tarquinio Evex Analytical 857 State road Princeton, NJ 08540
----------
I've been ignoring the microscopy meeting that is coming to Cleveland and suddenly my boss said to me today, "hey, we should think about going to this thing next week". We work in Cleveland so thats not a problem. Could someone send me information like schedules and fees and seminars and classes, etc, etc, etc. Thanks. Mark Darus
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JOB OPENING ANNOUNCEMENT!!!!!!!!!!!
Intel Corporation currently has an open position for a SIMS technician at its Santa Clara site's Materials Technology department. The SIMS group supports the Santa Clara site's development and fabrication facilities that produces state of the art microprocessors. The Materials Technology department's scope encompasses the whole process from silicon to a packaged device that includes process trouble-shooting, transfer and equipment qualifications, device failure analysis/fault isolation and new materials development to name a few. The department has a wide variety of state of the art instrumentation, including: SIMS, Auger, TEM, SEM/EDS, FIB, AFM, XRF, ICP-MS, FTIR, Raman, Pico and Nano Indentation.
Job Description:
The technician opening involves second shift operation of Quadrupole (Atomica) and Magnetic Sector (Cameca IMS- 6F) SIMS tools. SIMS is an Ultra High Vacuum analytical tool which uses ion beam sputtering combined with mass spectrometry detection to analyze dopant and contamination levels and distributions in patterned and unpatterned wafers. Duties will include data collection, data processing, report writing, interfacing with other engineers and customers, communicating results, and workflow duties to keep the lab running smoothly. The successful applicant must be self-motivated and capable of working with minimal supervision.
Qualification: An AA or BS degree with Engineering or physical science background or equivalent experience is required. Ability to work in a team oriented environment and good communication skills are critical. Experience with analytical equipment or Ultra High Vacuum or Vacuum systems is desired. Experience with SIMS and knowledge of Semiconductor processing methods is a definite plus.
Intel's industry leading total compensation package includes a competitive salary, stock options, annual employee bonus plan, bi-annual employee cash bonus payout, periodic paid sabbatical leave, and retirement plan. Intel is an equal opportunity employer.
Send resumes to or for more information contact:
Gabi Neubauer or Jerry Hunter Intel Corporation 2200 Mission College Blvd., M/S: SC2-24 Santa Clara, CA 95052
Phone: (408) 765-2241 or 765-2316 FAX: (408) 765-2393
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I've been ignoring the microscopy meeting that is coming to Cleveland and suddenly my boss said to me today, "hey, we should think about going to this thing next week". We work in Cleveland so thats not a problem. Could someone send me information like schedules and fees and seminars and classes, etc, etc, etc. Thanks. Mark Darus
We also use mag calibration tolerances of + or - 5%, and our company is ISO 9001 registered. However, I do not believe that ISO requires any specific tolerances for instrumentation; it simply requires that you determine what they are and conform to them. It is my belief that the "acceptable" tolerance is primarily dependent upon two things:
1) What can the instrument effectively yield? 2) What are your requirements? (i.e. How does the stated tolerance affect the quality of your data, and how critical is that?)
Just my thoughts on the topic.
Regards,
Bob ******************************* Bob Citron Chiron Vision Claremont, CA USA (909)399-1311 Bob_Citron-at-cc.chiron.com ******************************* ______________________________ Reply Separator _________________________________
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We have been using calibration tolerances of + or - 5% for both our magnification and spectrometer calibrations for quite some time now. Apparently these values have been established through supplier and user inputs quite some time ago. I am curious as to whether these values are consistent with those currently used or whether they are too loose. We are ISO 9002 registered and quality measures are very much a concern. TIA
Karpura Kommineni wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello everyone, } I am looking for a list of vendors of used light microscopes and parts. I } would like to have their phone number or other information that will help me } contact them. } You may reply directly to me. } Thank you } Sincerely } Karpura } kkommine-at-mbl.edu } Marine Biological Laboratory } Woods Hole Here are three vendors for used and new microscopes:
M2 Associates, Don Malatesta, 408-735-0495 McBain, 818-998-2702 Bender Associates, 602-820-0900
I hope these help.
Gary Liechty Allied High Tech Products, Inc. 2376 E. Pacifica Pl Rancho Dominguez, Ca. 90220 310-625-2466
I weep to hear that dihydrogen monoxide may be classified as a hazardous substance by the government authorities. Still, in some of our laboratories I have seen bottles of sand labled as hazardous material, presumably because they were labeled 'silica sand', and somehow or the othe anything with silicon or silica in it is considered taboo. Luckily, both are highly stable substances and can undoubtedly withstand the assult. However, I wonder when they'll start attacking hydrated hydronium hydroxide - a fearful sounding entity indeed, but one that is possibly more subject to having its reputation damaged than the others. If it were banned, we'd be in considerable trouble , indeed!
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
This subject is most distressing considering the material, also known as oxygen dihydride, has become so invasive in our daily routines. As many of you may be aware this materials the major ingredient in such pleasures as Guineas, Whatney's, John Courage, Old Speckled Hen,.... Need I say more?
Heath officials also recommend ingestion of at least 6 containers of DHMO/ODH daily - what are we to do?
Have a great day!! And thanks Mel for the insertion of a little humor!
Bob Craig
} ---------- } From: Melvyn Dickson[SMTP:M.Dickson-at-unsw.edu.au] } Sent: Thursday, August 07, 1997 1:01AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: New Toxic Hazard Shock } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Some eagle-eyed environmentalists have discovered yet another Government conspiracy to conceal the true harmful nature of a chemical which commonly contaminates the environment with dire results.
Full details of this biohazard, dhihydrogen monoxide, can be found at the following website:-
Steve Chapman's observations regarding SEM calibration are more than generous. Several years ago we did a calibration series on one of our SEM's using the Geller Standard and found that even though the instrument was well calibrated by the service engineer from the manufacturer - at the parameters that they specify for calibration - It was only valid for those conditions.
Any change in working distance, accelerating potential, beam current, etc., dramatically changed the validity of the calibrated values. For example, changing the working distance from 10mm to 30mm changed the magnification readout 20% from the actual standard values!!
These are things that are not usually mentioned or taught when dealing with instrument operation. A good point for any operator to be aware of when reporting "measured" values using a SEM.
As Steve also mentioned - forget about non-planar samples, or stage tilt.
Bob Craig OSRAM SYLVANIA Products inc. Lighting Research Center Beverly, MA 01915
} ---------- } From: Steve Chapman[SMTP:PROTRAIN-at-CompuServe.COM] } Sent: Wednesday, August 06, 1997 5:58PM } To: Wayne England; Msa link } Subject: Calibration tolerances } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to know what software is available commercially AND will run on a PC for doing optical sectioning by digital deconvolution. Presumably, such software would be supplied with parameters such as x,y,z resolution, wavelength, N.A., etc.
David P. Bazett-Jones, Ph.D.
Professor Departments of Anatomy and Medical Biochemistry The University of Calgary 3330 Hospital Dr Calgary, AB T2N 4N1 Canada TEL: 403 220-3025, FAX: 403 270-0737 email:bazett-at-acs.ucalgary.ca
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Karpura Kommineni wrote: } } } Hello everyone, } I am looking for a list of vendors of used light microscopes and parts. I
-- -------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
"Bother!" said Pooh as he was assimilated by the Borg "Bother!" said the Borg as they assimilated Pooh "Time for a little something" said the Borg "Oh dear!" squeaked Piglet while being assimilated by the Borg "Kanga is not a Borg identifier" squeaked the Borg
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The notice which follows was posted in March. Unfortunately the person to whom I offered the job has dropped out at the last minute. This job is available right now. If you are interested please contact me immediately. I will be at MSA in Cleveland next week - you can talk to me there or send me an e-mail message as soon as possible.
Microscopy and Computing
I am looking for a person to work on an exciting new project.
The appointment could be at the post-doctoral level or at other levels according to the background of the person appointed. In any case the post will be for three years.
The job is at the Materials Research Laboratory of the University of Illinois at Urbana.
The project is a joint enterprise involving Argonne National Lab, Oak Ridge National Lab, the Lawrence Berkeley Lab and NIST as well as the University of Illinois. The Project has the aim of developing a new kind of environment for electron microscopy and related techniques, in which the instruments can be operated remotely with the same effectiveness as they can be operated in the instrument room. More details of the project can be found at http://tpm.amc.anl.gov/MMC MMC is the abbreviation of the project name.
I am looking for someone who has familiarity with electron microscopy (preferably TEM) or a closely related technique - and who has well developed interests and experience in computing, particularly the interfacing of instruments for computer control and/or the networking of images.
Will any one interested please contact me right away. We would like the job to be started as soon as possible. ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
EMAG '97 Conference and trade exhibition ----------------------------------------
The biennial EMAG conference is being held in the Cavendish Laboratory, Cambridge from Tuesday 2 to Friday 5 September 1997. The principal aim of the conference is to promote and discuss recent advances in electron microscopy and related analytical techniques. Accompanying the conference is a major international trade exhibition and at present there are still a small number of stands available to potential exhibitors.
Conference sessions will be held on: microanalysis, semiconductors and superconductors, high resolution electron microscopy, ceramics/interfaces, electron crystallography, EELS, materials analysis, new instrumentation, advanced scanning probe techniques, microscopy of catalysis, intermetallics and advanced SEM and surface science.
For information on the conference see http://www.iop.org/IOP/Confs/EMAG/
For information on the exhibition or to book a stand at the exhibition see http://www-hrem.msm.cam.ac.uk/emag97/ or contact Chris Boothroyd, cbb4-at-cam.ac.uk Tel: +44 1223 334564 Fax: +44 1223 334567
At 08:36 AM 8/7/97 -0400, you wrote: -=-snip-=- } Any change in working distance, accelerating potential, beam current, } etc., dramatically changed the validity of the calibrated values. For } example, changing the working distance from 10mm to 30mm changed the } magnification readout 20% from the actual standard values!! } } These are things that are not usually mentioned or taught when dealing } with instrument operation. A good point for any operator to be aware of } when reporting "measured" values using a SEM. } } As Steve also mentioned - forget about non-planar samples, or stage } tilt. -=-snip-=-
Hello List,
Many modern SEMs take into account HT and working distance and adjust the mag accordingly (I know ours have for at least 10 years). Any given magnification on these scopes is usually +/- 5% of the displayed mag although there may be more than 5% difference compared to another mag.
Dave Harrison Site Manager JEOL USA, INC
"The trouble with America isn't that the poetry of life has turned to prose, but that it has turned to advertising copy." Louis Kronenberger
Does anyone have resin embedded, encephalitis infected tissue that they would be willing to share? After our son's illness, my husband and I started a grassroots organization (which appears in our email). Consequently, I decided to postpone med-school for two years to do some serious research. My dilemma is that I am attending a State funded school that will not permit me to generate my own tissue samples. The reasons: Animal Rights Committee regulations, the nature of the pathogen, and, of course, funding. The University is willing to back the endeavor--once the resin blocks are in hand. Any information would be greatly appreciated.
Sincerely, Debra Caires San Jose State University Biology Department One Washington Square San Jose, CA 95192 (408) 298-2060
As Dave Harrison states, all modern SEMs compensate for variables such as Working Distance, KV, Spot Size, etc.
Older SEMS had Mag readouts with only most significant digits and large steps between values.
Newer SEMS have more accurate mag readouts with smaller steps such as 1000, 1001, 1002, etc.
Many modern instruments have a Magnification Calibration procedure which adjusts the Mag to the Calibration sample used. If this method is used, and then the Mag is measured with the same sample, errors of less than 1% are possible. However, if you calibrate with one sample and then measure with another, then the Mag is only as good as the accuracy of the samples.
One of the funniest things that I have seen is safety data for water in one of these books of chemical hazards. It had phrases like: if splashed in eyes rinse with plenty of WATER, and in case of a spillage wash down drain with plenty of WATER!!
It makes you wonder sometimes.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
On Fri, 8 Aug 1997, Ian MacLaren wrote: } } One of the funniest things that I have seen is safety data for water in one } of these books of chemical hazards. It had phrases like: if splashed in } eyes rinse with plenty of WATER, and in case of a spillage wash down drain } with plenty of WATER!!
Ian. here is the copy of that text, sent on this list a few month ago:
Hello all
This is a slightly facetious input re. safety data sheets!
In the UK there is a very useful pair of publications from BDH, a chemicla supply company (used to be known as British Drug House, I believe). These are collcted data sheets. Yes folks, there is one for water. Here are a few salient points which all users of the substance should bear in mind at all times:
1. colourless liquid - maybe that implies it is rare and difficult to see if dropped! 2. Against solubility in water: miscible in all proportions. 3. fire and explosion hazard: not applicable (thats a relief - although if it caught fire I suppose one could foolishly attempt to extinguish with more water?). 4. Health hazard: no significant hazard expected, may be irritating to the eyes. 5. Toxicity: no data. 6. Carcinogenicity: no evidence of carcinogenic properties. 7. First aid - eyes: irrigate thoroughly with water(!) 8. First aid - lungs: remove from exposure. 9. First aid - skin: wash off thoroughly with soap and water (work that one out!). 10. First aid - mouth: wash out thoroughly with water. In severe cases obtain medical attention(!) 11.Reactive hazards: violent reaction with acyl halides, alkali and alkaline earth metals .... 12. Spillage disposal: wear appropriate protective clothing - listed are gloves, goggles, face shield and protective apron. Luckily, respirators are not needed.
I have an MSDS for soap. Skin contact requires washing with soap and water. However, it is not clear if you can use the SAME soap or if you need to use a DIFFERENT soap. Since this problem is unresolved, I just hope and pray I never get any soap on me. What would I do?
Many :-)
} ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com}
Let me describe the procedure we used to determine variations in magnification values, etc.
The instrument was calibrated against the Geller Standard by the manufacturers service engineer using their procedures. Magnification, x vs y, image squareness, etc., - all within =B1 1%. In all cases the = beam was orthogonal to the standard surface. Changing the working distance from 10mm to 30mm, refocussing on the standard, removing hysteresis from the column (i.e., "ringing" the lenses), adjusting the magnification to the same value as at calibration, recording an image, measuring the same pair of lines on the standard and comparing these values with the original calibration image resulted in an error of -20%. We also did this procedure at several intermediate working distances and found the variation was not linear with working distance as well.
Going to shorter working distances had a positive effect, but no where near as large as with longer working distances, i.e., the rate of change in off-calibration per mm change in working distance was less for shorter working distances.
Nothing was changed except the working distance and the difference in lens and scan coil current required to produce crossover on the standard at the lower position. The beam current was not adjusted to compensate for the difference in lens current (which normally would not be done unless doing "quant" EDS).
I know nothing about, nor do I really care about, what compensating circuits are supposed to do, etc., just that when parameters are changed one must be aware that readouts may not be telling the "truth and nothing but the truth."
It is my opinion that many SEM operators at not aware of these little vagaries and should be made aware of them and realize that those neat little digital readouts might not be accurate under conditions different from the instruments calibration parameters.
Bob Craig OSRAM SYLVANIA Products Inc. Lighting Research Center Beverly, MA 01915
Hi, As organiser of the forthcoming First Electronic Analytical Chemistry Conference, I would like to invite someone well know in the field to present an interesting talk on Microscopy. Since this is not my field I would like suggestions as to who to approach with an invitation.
The conference will take place over the internet in early November and will take the form of previous electronic conferences which were very successful.
ECCC1 & ECCC2 First and Second Electronic Computational Chemistry http://hackberry.chem.niu.edu:70/0/ECCC/homepage.html
ECHET96 Electronic Conference on Heterocyclic Chemistry http://www.ch.ic.ac.uk/ectoc/ehet96/
During the conference interaction, presentations and discussions will take place via the Internet using a Java-based virtual conference centre, WWW-based discussion forums and an electronic mailing list.
The Iowa Microscopy Society will hold its annual symposium on Thursday, September 25, 1997 at the Eckstein Medical Research Building on the University of Iowa campus in Iowa City.
TENTATIVE MEETING AGENDA September 25th
7:30-9:00am Registration Poster and display set-up
8:00 Welcoming Remarks
8:20 "Morphological Characterization of the Metastatic Mary Hendrix Department of Anatomy, University of Iowa
8:50 "HIV-Induced T-cell Syncytia, In Vivo and In Vitro, are Motile, Invasive and Destructive" Karla Daniels Department of Biological Sciences, University of Iowa
9:35 "To be announced" Charles Gross Department of Microbiology, University of Iowa
10:05 BREAK
10:30 "Fractility: Correlation of Images with Transport Characteristis of Exchange Polymer Composites" Johna Leddy Department of Chemistry, University of Iowa
11:00 "Applications of Multiphoton Excitation Imaging." Victoria Centonze-Frohlich IMR, University of Wisconsin
12:00 LUNCH
1:00 "Same Cell Correlative Video Enhanced Light and 3-D Electron Microscopic Studies of Mitosis in Vertebrate Cells" Conley L. Rieder NIH
2:00 "The Effect of Mutation of COMP on Calcium Deposition in Chondrocytes" Jeff Stevens Department of Orthopedic Surgery, University of Iowa
2:30 BREAK
3:00 "BCL-2 Expression Alters Targeting of Viral Products in Insect Cells" David Murhammer Department of Chemical Engineering, University of Iowa
3:30 "Advances in Ultra-Small Gold Probes in Light and Electron Immunocytochemistry and In-Situ Hybridization" Peter Van de Plas AURION, The Netherlands
4:30 "Fluorescent Annual Banding in Speleothems, Applied to Paleoclimatology" Christopher Shorey Departmnt of Geology, University of Iowa
5:00 IMS Business Meeting
5:15 RECEPTION
Symposium costs are:
Preregistration: Meeting day registration: regular-$8.00 regular-$12:00 student-$5.00 student-$7.00
For a registration form or further details please contact Kathy Walters (addresses below).
IMMUNOCYTOCHEMISTRY WORKSHOP
In addition to the symposium, a workshop is offered on the following Friday and Saturday (September 26th and 27th). Peter van de Plas will provide a general lecture of immunocytochemical techniques and hands on instruction. For more details please visit the CMRF website at:
http://www.uiowa.edu/~cemrf
POSTER COMPETITION
Three cash prizes in two catagories will be awarded to outstanding participents. Applications for submitting posters to the meeting are now being mailed. If you would like an application, or would like to be added to our mailing list, please contact Kathy Walters at the address below.
Kathy Walters / / Research Assistant III / /\ Center for Microscopy Research / /\ \ University of Iowa /_/ \ \ 85 EMRB ____ ((O)) Iowa City, Iowa 52242 | | / / || / / email: kwalters-at-emiris.iaf.uiowa.edu ----------- fax: (319)335-8049 ------------- www: http://www.uiowa.edu/~cemrf
I would be grateful if someone could point me to a reference on the imunno electron microscopic localization of carbonic anyhdrase in mamallian tissue. No hurry since I am leaving for Cleveland on Sunday. See you all there.
Greg Erdos ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Can anyone help me find a supplier for EMHOPC (ethyl-1-methyl-4-hydroxy-oxo-3-pyrroline-3-carboxylate)? It is a ferric ion chelating agent that has been reported to give permanent staining of ATPase activity in skeletal muscle fibers, ref: Hawcroft DM ,Ball MT A new chelating method for determining ATPase activity in skeletal muscle. Biotechnic & Histochem 1996; 71 (2): 88-91. I have tried contacting the authors by mail, but no reply. Perhaps some of my fellow Brits can help, as Drs Hawcroft and Ball are employed in the Department of Pharmaceutical Sciences, De Monfort University, Leicester, England. Thanks, in advance, for any assistance. Ronnie Houston Director Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children Dallas, TX 75219 USA
Applied Precision 8505 SE 68th Street Mercer Island, WA 98040
Phone: 206-236-0704 Fax: 206-232-4184
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
-------- REPLY, Original message follows --------
I run the Electron Microprobe Lab at Arizona State University, where we have A JEOL 8600 probe with (Noran Tracor) 5500 Series II EDS system and 5600 PAC automation system. The Vista image analysis software works fairly well - however, Noran did not engineer the capability to export the images to the outside world in a real-world format. We have found a way around this problem by using the } XI 2 Terminal Emulator supplied by Noran. The image has to be transmitted as a TEXT file, and the receiving PC using SmartTerm receives it as a text file. It takes five minutes to transfer a 512x512 gray-scale image, such as SEI and BEI. The PC disk then has to be transferred to a MAC to convert it to a standard TIFF file through the NIM Image program. While it does work, it is understandably slow and a real pain.
The problem that I am trying to resolve now is figuring out how to transfer EDS x-ray dot maps. The closest I've coming to succedding is to collect the map for a single element, convert it to a binary image, use } XI 2 to transmit as a binary image to The PC using SmartTerm to receive it as a binary image. I get an image on the MAC, but it is barely visible. The dots are very faint gray instead of white, and the background is an intermediate to light gray instead of black. I had stored the binary image in Binary 1, and left the other Binary bins blank. I had also tried copying the faint image in Binary 1 into Binary 6 thru 7, so that all bins contained the same image. No Luck. Finally, I tried transmitting the lone Binary image in Binary 1 and transmitting it as a binary image to SmartTerm as a TEXT file. Again, no luck.
I seem to have run out of ideas. I have called Noran, but they had no suggestions. Todd Jarlsberg had been working on this problem and seemed to have a pretty good handle on it, but left Noran a couple of years ago.
Has anyone out there been down this same road, and if so have you come up with a solution, or at least have any suggestions. We would prefer if at all possible to resolve this problem without having to add upgrades to the system, money being tight as it is these days, but that may be unavoidable.
Thank you for your ideas and efforts.
Jim Clark e-mail: jclark-at-asu.edu Probe Lab: 602/965-61720
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } JOB OPENING ANNOUNCEMENT!!!!!!!!!!! } } Intel Corporation currently has an open position for a SIMS technician at its } Santa Clara site's Materials Technology department. The SIMS group supports } the Santa Clara site's development and fabrication facilities that produces } state of the art microprocessors. The Materials Technology department's scope } encompasses the whole process from silicon to a packaged device that includes } process trouble-shooting, transfer and equipment qualifications, device failure } analysis/fault isolation and new materials development to name a few. The } department has a wide variety of state of the art instrumentation, including: } SIMS, Auger, TEM, SEM/EDS, FIB, AFM, XRF, ICP-MS, FTIR, Raman, Pico and Nano } Indentation. } } Job Description: } } The technician opening involves second shift operation of Quadrupole (Atomica) } and Magnetic Sector (Cameca IMS- 6F) SIMS tools. SIMS is an Ultra High Vacuum } analytical tool which uses ion beam sputtering combined with mass spectrometry } detection to analyze dopant and contamination levels and distributions in } patterned and unpatterned wafers. Duties will include data collection, data } processing, report writing, interfacing with other engineers and customers, } communicating results, and workflow duties to keep the lab running smoothly. } The successful applicant must be self-motivated and capable of working with } minimal supervision. } } Qualification: An AA or BS degree with Engineering or physical science } background or equivalent experience is required. Ability to work in a team } oriented environment and good communication skills are critical. Experience } with analytical equipment or Ultra High Vacuum or Vacuum systems is desired. } Experience with SIMS and knowledge of Semiconductor processing methods is a } definite plus. } } } Intel's industry leading total compensation package includes a competitive } salary, stock options, annual employee bonus plan, bi-annual employee cash } bonus payout, periodic paid sabbatical leave, and retirement plan. Intel is } an equal opportunity employer. } } Send resumes to or for more information contact: } } Gabi Neubauer or Jerry Hunter } Intel Corporation } 2200 Mission College Blvd., M/S: SC2-24 } Santa Clara, CA 95052 } } Phone: (408) 765-2241 or 765-2316 } FAX: (408) 765-2393 }
A colleague is seeking a means for tranferring images recorded on 2" video floppies to more standard digital format. These video images were generated/stored with a Hitachi VX-100A Video Floppy System. However, we don't have the system here; just the disks. In fact, we can't even find information on this system on Hitachi's website.
So my questions are: Has anyone ever heard of this Hitachi video floppy system? What is required to read the disks? Most importantly, does anyone know of a local (Minnesota) system that could read these disks and transfer them to a computer? Even transfer to VHS format could be useful, if nothing else is available.
Thanks as always for your help and suggestions! Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
I wish to hear from fellow users of the JEOL 6400F, if there are any out there. Specifically, I am curious about your experiences running the system at low kV, 600V to 3kV accelerating voltage. Have you experienced any image instabilities? Running in "super rapid", I have seen the entire image shift about 1/4 to 1/2 inch and then return to the original position. In photos, this shows up as a band in the image that is shifted sideways. Another anomaly I have seen is the brightness jumping up and down. In photos this shows up as brighter and darker bands horizontally across the photo. If you have seen these effects, has there been any solution or explanation? (Other than it is being caused by fields, probably from Alfa Centari! :-) )
Used Microscopes can be obtained via several sources on our Services Page. You could also place a request for individual mcroscopists who might be selling off their instruments on our Forum pages... all at: -
http://www.microscopy-uk.org.uk
These pages, and the site, predominantly cater for amateur (optical) microscopists... but of course - everyone is welcome!
I am not familiar with the Tracor/Noran format, so may may wish to contact John Mansfield at U-Mich. He has a Tracor and has done networking with it to other platforms. I suppose he knows of a method or utility for handling maps. There are converter utilities for many things thru the file archives. But again, John will have to point you to the right files. You can start with the URL ftp://www.amc.anl.gov/AMC-3/ANLSoftwareLibrary/ and go from there.
I would hope you can pick off the map parts and transfer them over as an image without going binary first. You would probably need to normalize the grayscale after transfer. Our Link stores the count as the grayscale so that we only hit 255 on very long maps. Most of our maps run from 0 to less than 40. Many packages can remap that to a 0 to 255 range so that the trends are visible.
Good hunting.
At 09:16 AM 8/8/97 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am trying to assist a research group interested in looking at nanoparticles. Some are made of silver iodide and others are made of cadmium sulfide, sometimes with a little zinc. They are supposed to be about 5 micrometers in size. They do the synthesis and some sorts of analysis, infrared spectra, I think. They come to my TEM to check on the size.
Well, I am having a hard time seeing them. I tried a drop of suspended particles on a formvar coated grid, a technique that usually works OK. I could hardly see any particles in the TEM. I saw a few particles that might have been what they are looking for, but they were few and far between. I suggested that the concentration might be low, they said, no, should be lots in there.
Without going into lots of details here, I promised them I would try to find out how to look at their particles. I said I would try a question to the list. Soooo, if you think of any helpful hints send them my way.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
You know those 5 micrometer particles I couldn't see? Well, they are really supposed to be 5 nannometers (duh). You see I was so excited about getting ready to go to Cleveland that I lost my head. Hopefully nobody will notice this dumb mistake until after I get back from the meeting!
See (saw?) you there.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Dear Jon, With particles that huge, the suspension should be cloudy if it is of any concentration. In fact, you should be able to look at a dried suspension on polished graphite in the SEM and check the composition by EDX. Just because their recipe says it should produce these particles, doesn't mean it necessarily will. I have been looking for spheres of Ni, 50 nanometers diameter, that the Chemical Engineering Department has been trying to make through two Master's theses. So far we have seen maybe five. Can you use a carbon film? It is clearer. With those heavy elements, imaging a dried suspension on a formvar should show the particles clearly at 100kV on the TEM. Do you have EDX on your TEM? I have looked at silver chloride crystals, silver spheres 30 nm. in diameter and the aforementioned nickel spheres. The main problem is the other dissolved solids in the suspension obscuring the whole grid, in which case the whole grid is dark and blurry. You wrote:
} Help! } } I am trying to assist a research group interested in looking at } nanoparticles. Some are made of silver iodide and others are made of } cadmium sulfide, sometimes with a little zinc. They are supposed to be } about 5 micrometers in size. They do the synthesis and some sorts of } analysis, infrared spectra, I think. They come to my TEM to check on the } size. } } Well, I am having a hard time seeing them. I tried a drop of suspended } particles on a formvar coated grid, a technique that usually works OK. I } could hardly see any particles in the TEM. I saw a few particles that might } have been what they are looking for, but they were few and far between. I } suggested that the concentration might be low, they said, no, should be } lots in there. } } Without going into lots of details here, I promised them I would try to } find out how to look at their particles. I said I would try a question to } the list. Soooo, if you think of any helpful hints send them my way. } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} We are shopping for a TV rate camera to interface to our Hitachi H-600 TEM } via a 35 mm camera port (scope is equipped with STEM). We plan to use } this for teaching and when more than one person (i.e. operator and } researchers) are working at the scope, not for acquisition of high-quality } digital images. To the best of my knowledge these systems work as } follows: a small phosphor screen will be moved into the beam path, a } camera is focused on this screen and the image is displayed on a } small B&W monitor. I have found two vendors so far (Fullam and Gatan), } but our purchasing department wants more. If you know of any other } vendors for this kind of equipment - or are vendors of it - please reply } directly to me by e-mail. } } Thanks in advance for any replies. } } Heather Owen
If you receive, or can get hold of, a copy of Microscopy & Analysis, there are adverts from the main companies plus a buyers guide - if you use the buyers guide, you'll get all the info for a specific product area sent to you.
I have an interest here, as Technical Editor.
You also need to distinguish between lens coupled systems, which is what you describe, and directly coupled system, where the camera itself is placed inside the vacuum of the TEM. A transmission phosphor (YAG) is used, which is coupled by a fibre optic plate to the camera (usually a CCD).
Generally, you will find that directly couple systems are more efficient a collecting the light from the phosphor than lens coupled systems.
Regards,
-- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis M&A web site - 17, Rocks Park Road http://www.microrgc.demon.co.uk Uckfield, E. Sussex email: LPS-at-teknesis.demon.co.uk TN22 2AT Phone: +44 (0)1825 766911 United Kingdom Fax: +44 (0)1825 766911
Hello - I don't know about silver iodide, but I've been assisting a bit in= using TEM to look at 2-5 nm diameter CdS particles stablized using either= low molecular weight ligands, or block copolymers. They show a lot less BF= contrast than gold, say. If there are a lot of them, close to Gaussian= focus the overall texture looks more like amorphous carbon at high defocus= than a neat array of distinct little round balls (and it looks like a= complete mess at high defocus, especially given all the organic stuff in= there).=20 So, assuming you aren't using EDX to check for Cd, and that you are= not capable of imaging CdS lattice fringes, then it might be that you're= just not seeing the trees for the wood. In that case, try diluting the= suspension and picking the particles up on a thin carbon film (rather than= formvar). It's also an idea to use a holey carbon film - then you should be= able to see some of the particles hanging over the edges of the holes, even= if they are not showing up well against the carbon.=20 In my experience, if the light absorbtion data say you have a lot of= little particles in suspension, then with the droplet method you generally= end up with a lot of little particles on the carbon film (especially with= CdS, which seems to be everybody's favorite system). In such cases we go= straight to lattice imaging on an EM 430/LaB6 to get the particle sizes (so= that it doesn't particularly matter if the low resolution BF images don't= show a great deal). If the nanoparticles all decide to clump together and= float away during the drying step, then you can usually see that happening= in the optical microscope, and freeze drying is in order. If they're= already clumped together then the suspension will be cloudy and the= chemists need to get their act together. =20
Pr=E9nom Nom: Christopher John George Plummer Institut: Laboratoire de Polym=E8res, D=E9partement des Mat=E9riaux EPFL: Ecole Polytechnique F=E9d=E9rale de Lausanne CH-1015 Lausanne t=E9l: (+41 21) 693 28 56 email: christopher.plummer-at-lp.dmx.epfl.ch
Characterization of 5nm-sized AgI and Cd(Zn)S particles requires TEM and STEM techniques with an appropriate resolution and a higher electron beam intensity, which can be readily obtained with microscopes equipped with LaB6 or field emission guns. It may need also preparing thinner supports than conventional formvar films. Better results one can achieve by the use of amorphous carbon films and carbon holey films up to 2nm in thickness. Here are some references for the reproducible protocol for making such films:
Procedures in Electron Microscopy /Eds. A.W.Robards and A.J.Wilson. J.Wiley&Sons, Chichester, 1993,pp.4:6.15-6.22. W.Baumeister and J.Seredynsky/Micron, 1976, v.7, pp.49-54.
Some problems may cause aggregation and coalescence of particles during deposition on film-supports. Therefore particles in a suspension of an appropriate concentration should be stabilized by adding of a protective polymer such as polyphosphate or by suitable complexing ligands.
In my experience, electron beam-induced decomposition and related phenomena especially in the case of AgI nanoparticles resulting in quick reduction to silver with elimination of halide, melting, evaporation and recrystallization of particles may significantly hamper studies. For such observations it would be necessary to use a cryo-cooling at liquid nitrogen temperature. One can also reduce damage, modification and drift effects by working fast and/or using minimal electron doses.
Regards,
Vladimir Oleshko ********************************************************** V.P. Oleshko, Ph. D. e-mail:oleshko-at-uia.ua.ac.be Micro-and Trace Analysis Centre Tel.:+32-3-820.23.64 Chemistry Department FAX :+32-3-820.23.76 University of Antwerp (UIA) Universiteitsplein 1 Antwerpen-Wilrijk B-2610 Belgium ***********************************************************
On Fri, 8 Aug 1997, Jon Krupp wrote: } Help! } } I am trying to assist a research group interested in looking at } nanoparticles. Some are made of silver iodide and others are made of } cadmium sulfide, sometimes with a little zinc. They are supposed to be } about 5 micrometers in size. They do the synthesis and some sorts of } analysis, infrared spectra, I think. They come to my TEM to check on the } size. } } Well, I am having a hard time seeing them. I tried a drop of suspended } particles on a formvar coated grid, a technique that usually works OK. I } could hardly see any particles in the TEM. I saw a few particles that might } have been what they are looking for, but they were few and far between. I } suggested that the concentration might be low, they said, no, should be } lots in there. } } Without going into lots of details here, I promised them I would try to } find out how to look at their particles. I said I would try a question to } the list. Soooo, if you think of any helpful hints send them my way. } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } } }
{bold} {color} {param} FFFF,0000,0000 {/param} {bigger} {bigger} PHILIPS EM201 FOR SALE
{/bigger} {/bigger} {/color} {/bold} A Philips EM 201 transmission electron microscope in excellent condition is available for immediate sale. The instrument had been used basically by one person, and is covered by service contract. A new plate camera was installed last year. Asking price is $3500. Crating and shipping charges would be buyer's responsibility. If interested, please contact:
Dr. Cornelia Farnum
Anatomy Department, College of Veterinary Medicine
The University of Glasgow and The Queen's University of Belfast are seeking 2 Postdoctoral Research Assistants for the following=20 research project.
THE USE OF XANES AND ELNES FOR THE CHARACTERISATION OF STABILISED ZIRCONIA= =20
The University of Glasgow and the Queen's University, Belfast have been awarded a major EPSRC grant to investigate and develop the use of near edge fine structure in X-ray absorption spectroscopy (XANES) and electron energy-loss spectroscopy (ELNES) for the characterisation of commercially important zirconia-based materials. This multidisciplinary project involves researchers in four universities, as well as researchers at the Daresbury Laboratory and in two UK companies, MEL Chemicals Ltd and Johnson Matthey= Ltd.
We are seeking two outstanding postdoctoral research assistants (PDRA) to work on this project. PDRA1 will be based in Glasgow for 30 months and will carry out the experimental work using the analytical electron microscopy facilities at Glasgow and the synchrotron facilities at Daresbury. PDRA2 will be based in Belfast for 12 months and in Glasgow for the following 18 months and will develop and apply theoretical models for calculation of ELNES & XANES in defective zirconia materials. Both posts require candidates with a practical approach to problem solving and should appeal to those with a background in solid state chemistry/condensed matter physics/materials science. For PDRA1, experience in electron microscopy, especially electron energy loss spectroscopy, would be an advantage. For PDRA2, experience with first principles band structure calculations is essential and a background in the theoretical interpretation of spectroscopic techniques such as ELNES and XANES would be highly desirable, as would a knowledge of many-body physics.
Initial salary will be on the RA1A scale up to =A316,927 per annum,= depending on qualifications and experience. Applicants should send two copies of their CV along with the names and addresses of two referees to Dr David McComb, Department of Chemistry, University of Glasgow, Glasgow G12 8QQ, UK.
Informal enquiries and requests for further details can be made to Dr Alan Craven (0141 330 5892, a.craven-at-physics.gla.ac.uk), Dr David McComb (0141 330 4486, davidm-at-chem.gla.ac.uk) or Prof. Mike Finnis (01232 335330, M.Finnis-at-qub.ac.uk). Further details can also be obtained on the following websites; www.chem.gla.ac.uk, www.ssp.gla.ac.uk, titus.phy.qub.ac.uk.
The closing date for applications is 12 September, 1997. ----------------------------------------------------------------------------= ---- Dr David W McComb Department of Chemistry University of Glasgow Glasgow G12 8QQ U.K.
I once worked on solar-cell multilayers, and found that almost all of those compounds, including CdS, were very beam sensitive. They evaporate and leave nothing during observation, or re-deposite on the specimen and destroy any nice trasparent areas in ~10 sec. at 120kev. I believe you may be using a higher kev (200kev?) to see 5nm particles. It seems to me that the particles could be gone before you saw them.
If you just need to know the particle size, a very thin carbon coating on your specimen may help you out. Otherwise, you may send me the particles. This company accepts TEM/OM consulting services of particles, thin films and bulks of any materials.
Chao-Ying Ni, PhD Manager of Microscopy Dept. Batta Laboratories Inc. 6 Garfield Way Delaware Industrial Park Newark, DE 19713
On Fri, 8 Aug 1997, Jon Krupp wrote:
} Date: Fri, 8 Aug 1997 16:33:31 -0700 } From: Jon Krupp {jmkrupp-at-cats.ucsc.edu} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Imaging AgI & CdS nanoparticles } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Help! } } I am trying to assist a research group interested in looking at } nanoparticles. Some are made of silver iodide and others are made of } cadmium sulfide, sometimes with a little zinc. They are supposed to be } about 5 micrometers in size. They do the synthesis and some sorts of } analysis, infrared spectra, I think. They come to my TEM to check on the } size. } } Well, I am having a hard time seeing them. I tried a drop of suspended } particles on a formvar coated grid, a technique that usually works OK. I } could hardly see any particles in the TEM. I saw a few particles that might } have been what they are looking for, but they were few and far between. I } suggested that the concentration might be low, they said, no, should be } lots in there. } } Without going into lots of details here, I promised them I would try to } find out how to look at their particles. I said I would try a question to } the list. Soooo, if you think of any helpful hints send them my way. } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } } }
Darrell Miles asked about shifting images on a JEOL 6400F:
We had similar problems on our JEOL 6000F (sudden lateral movement of the image). This was eventually, after MUCH investigation by all concerned, traced to very low frequency ( } } 1hz) magnetic fields. In our case the probable source is the railway approx 250m away from the microscope (but we have not ruled out Alpha Centaurus!). We have measured magnetic intensities up to 4mG, running in X, Y and Z directions. Due to the low frequency (approximating DC fields) it is not possible to monitor these fields with the normal multiturn coil connected to an oscilloscope, which is why it took us so long to identify the problem. Field emission microscopes seem to be much more prone than tungsten filament microscopes to magnetic interference (the other two scanners in the immediate surroundings are tungsten filament machines, JEOL 840 and 5800LV) and are much less affected. Shielding is not effective for the attenuation of these low frequency fields. Our solution was the installation of field cancelling equipment - expensive, somewhat of a nuisance, but effective. Darrell's second problem could be due to sparking on the SE detector scintillator - one way of checking this is to electrically connect the aluminium coating of the scintillator disk to the scintillator holder (a VERY small dab of carbon paste usually does the trick). If the brightness variation is seen also with the BE detector, the source of the variation is of course elsewhere - possibly in the noise canceller??.
} I wish to hear from fellow users of the JEOL 6400F, if there are any out there. } Specifically, I am curious about your experiences running the system at low } kV, 600V to 3kV accelerating voltage. Have you experienced any image } instabilities? Running in "super rapid", I have seen the entire image shift } about 1/4 to 1/2 inch and then return to the original position. In photos, this } shows up as a band in the image that is shifted sideways. Another anomaly } I have seen is the brightness jumping up and down. In photos this shows } up as brighter and darker bands horizontally across the photo. If you have } seen these effects, has there been any solution or explanation? (Other than } it is being caused by fields, probably from Alfa Centari! :-) ) } } Thanks, } Darrell
Prof Jan Coetzee Head: Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/science/electron/emunit1.htm
I have a Matrox Meteor imaging board and would like to acquire RGB images via a JVC TK1070E CCD camera fitted to a Zeiss confocal microscope. Does anyone know of any shareware (or fairly cheap) image acquisition software that I could use to view and acquire images, preferably as TIFF image ?
Regards
Mark Auty DPC Moorepark Fermoy Co. Cork, Ireland mauty-at-dpc.teagasc.ie
I want to study the cell walls changes of surface cells during frying of potato. My idea is:
1st- colour the potato cells at the surface and observe it with a light surface microscope
2nd- fry the potato at 180 celsius
3rd- observe the same cells at the surface without any other treatment
The first step it is easy to do. I used safranin to colour the cell walls. But, when I fry the potato, the dye disappears.
This may sound ridiculus but, does anyone know of a dye or a treatment that may fix the dye to cell wall even after heating to temperatures of 180 celsius degrees ?
I had to look at nano particles of Ag before and I know what you have to deal with. What I found best was to a specimen of sputtered Ag on a carbon support grid to use as a standard. I then proceeded to set up for darkfield conditions using one of the rings produced by the standard. When I went back to the particles they lit up under the darkfield conditions and were easy to locate. You may try something similar with a crushed standard of AgI and CdS. Good luck.
Roberto Garcia EMF Manager Wright State University
I have warn you on the subject line. This is tentative answer for CTF correction in image reconstruction. If you are not interested in this subject, or even you are, this is going to be a long, boring email!
Thank all the people who give me clues or references! After the brief reading of some of those literature, I try to come up with answers to those questions I put up last time. If anything is wrong regarding these answers, it is my fault, not the person who gave me hints!
} What kinds of contrast exists in electron microscopic image } for weak-phase object? Is amplitude contrast the same as } aperture contrast?
For weak-phase object like thin biological specimen, there are basiclly two kinds of contrast: amplitude and phase contrast. Phase contrast is produced by the interference of the scattered electron wave with the unscattered or background electron wave. Amplitude contrast is produced by the loss of electrons which are scattered outside the objective aperture.
} What is the contribution of inelastic scattering on the image } contrast? How to account for it?
During inelastic scattering the incident electron loses energy in the specimen and produces a significant spread of wavelengths, so large in fact that the scattering of these electrons produce essentially a continuous background in the electron diffration pattern.
Zero-loss energy-filtered images of frozen-hydrated specimen have been shown to have higher contrast and improved structural resolution.
} How does partial spatial coherence and temporal coherence } influence the image and electron diffraction?
The coherence of electron source has an important role in the high resolution imaging of weak-phase objects. The partial temporal and spatial coherence both contribute as a multiplicative factors on the CTF, imposing a resolution limit by attenuating high order spatial frequencies. In the focal plane, the partial coherence broadens the diffraction spots.
} Is it always sufficient to use just first order approximation } ( the linear theory of phase and amplitude contrast image } formation ) for CTF consideration?
Yes, it works fine in most cases of biological samples.
} Is it true for low spatial frequencies ignoring the CTF was } better than compensating for phase contrast alone?
In this regime the amplitude contrast is most important. Phase contrast vanishes, and if you correct for this by the inverse of the phase CTF, the error will be considerable.
} I guess compensation for the CTF was necessary and sufficient } to accurately reconstruct molecular densities. Could anyone tell } me the current ways for accurate determination of CTF?
There are basiclly two ways to approach CTF correction. The first one is using diffractogram to obtain the Thon rings, from that to estimate the defocus values or even the ratio of amplitude contrast to phase contrast by the use of two different defocus settings.
Dr. Frank have introdued a second method which accounts for not only the amplitude and phase contrast components, but also all the envelope functions which attenuate the high resolution information and noise. Least-square refinement of the theoretical value against experiment data gives rise to the each parameters in the CTF. They even combine the CTF correction with the 3D reconstruction in a single step to reduce the error and improve resolution of final results.
A recent paper by Ichise described a phase spectra based method in the measurement of TEM parameters ( defocus, astigmatism and beam tilt misalignment ) and the image drift. The phase spectrum is the phase part of the cross-spectrum between two different images due to beam tilts, in which the cross-spectrum is defined as a Fourier transform of the cross-correlation function.
For 2D crystal, Dr. Henderson had set up the ways to accound for phase shift due to tilts and the refinement proticol in astigmatism correction. Spot-scan is supposed to reduce phase gradient during tilts. But for tomograph, I have no clue how to correct phase gradient.
In a recent paper, Dr. Fuller have pointed out the improvement of resolution in icosahedral virus reconstruction attributed not only the number of particles used but also the improved quality of the data from better microscope. And also stressed the importance of CTF correction in the final structure.
} By improving the different components in CTF, what is the optimal } contrast could be achieved in image?
This is mainly limited by the noise. Dr. Brink reported a contrast at 0.42 by spot-scan at 400 KV.
} Is it possible to put EM reconstruction on the same scale with } the structures derived from X-ray crystallography and NMR after } deliberate correction of CTF, solvent effects and differences between } atomic scattering factors for electron and X-ray?
Absolutely.
} To what extend we could use the insights obtained by building } models and comparison of the models with EM reconstruction?
From the limited knowledge of mine, there are several groups trying to gain insights from combining EM data with X-ray structures. Dr.Stewart reported the combining of capsid protein structures from X-ray into EM reconstruction, and discussed several issues on the combination. Dr. Baker have done a lot of work on the virus structural and functional studies based on the results of the known crystal structures and their EM reconstructions. Dr. Milligan et al. derived the mechanism of muscle contraction based on docking the myosin and actin crystal structures into cryo-EM reconstruction of muscle fibers. Dr. Frank combine the crystal structure of tRNA into ribosome 3D reconstruction to deduce the protein translation machanism. There might be tons of other work out there I just have not read yet, please forgive me!
References:
Books:
"Experimental high-resolution electron microscopy", Spence, John C.H., New York : Oxford University Press, 1988. "Transmission Electron Microscopy", Reimer, Ludwig, Berlin ; New York : Springer-Verlag, 1989. "High-resolution transmission electron microscopy and associated techniques" edited by Peter Buseck, John Cowley, and Leroy Eyring. New York : Oxford University Press, 1988.
Journal articles:
Erika J Mancini, Felix de Haas and Stephen D Fuller.(1997) High-resolution icosahedral reconstruction:fulfilling the promise of cryo-electron microscopy. Structure 5 : 741-750.
Zhu J; Penczek PA; Schrvder R; Frank J. (1997) Three-dimensional reconstruction with contrast transfer function correction from energy-filtered cryoelectron micrographs: procedure and application to the 70S Escherichia coli ribosome. J Struct Biol, 118: 197-219
Frank J; Penczek PA. (1995) On the correction of the contrast transfer function in biological electron microscopy. Optik, 98: 125-129
Wade R.H.; Frank J. (1977) Electron microscope transfer functions for partially coherent axial illumination and chromatic defocus spread. Optik, 49:81-92
Ichise N.; Baba N.; Nagashima H. (1997) The phase spectrum-based measurement of the TEM parameters. Ultramicroscopy. 68:181-200
Erickson H.P. & Klug, A. (1970) Measurement and compensation of defocusing and aberrations by Fourier processing of electron micrographs. Phil. Trans. Roy. Soc. Lond. B261, 105-118
Erickson, H.P. (1973) The fourier transform of an electron micrograph - first order and second order theory theory of image formation. In: Advance in optical and electron microscopy, 163-199
Smith, M.F. & Langmore, J.P. (1992) Quantitation of molecular densities by cryo-electron microscopy: Determination of the radial density distribution of tobacco mosaic virus. J. Mol. Biol. 226, 763-774
Schroder, R.R., Hofmann, W. & Metrenet, J.F. (1990) Zero-loss energy filtering as improved imaging mode in cryoelectronmicroscopy of frozen hydrated specimens. J. Struct. Biol. 105, 28-34
Stewart, P. L., S. D. Fuller and R. M. Burnett (1993) Difference imaging of adenovirus: Bridging the resolution gap between x-ray crystallography and electron microscopy. EMBO J. 12:2589-259.
Toyoshima, C. and N. Unwin (1988) Contrast transfer for frozen-hydrated specimens: determination from pairs of defocused images. Ultramicrosc. 25:279-292.
Toyoshima, C., K. Yonekura and H. Sasabe (1993) Contrast transfer for frozen-hydrated specimens II. Amplitude contrast at very low frequencies. Ultramicrosc. 48:165-176.
Brink, J. and W. Chiu (1991) Contrast analysis of cryo-images in N-paraffin recorded at 400kV out to 2.1E resolution. J. Microsc. 161:279-295.
Any comments or corrections will be heartly welcomed! By the way, I have put a copy of this at the end of my cryo-EM reconstruction homepage: " http://www.sb.fsu.edu/~jinghua/em.html "
Yves This may seem a trivial thing but is a source of constant annoyance when I am involved in COSHH (Control of Substances Hazardous to Health) risk assessments because it isn't just water but lots of others including NaCl, PBS, sucrose, glucose. It would be useful if suppliers of these data sheets could preface their hazard information with detail such as low risk unless ingested by the kilogram because sometimes you can very easily come across more hazardous materials with little known information which may seem more innocuous. Some people will then argue, according to the data, that the reagent is no more dangerous than sodium chloride solution.
Malcolm Haswell e.m unit University of Sunderland UK ----------
On Fri, 8 Aug 1997, Ian MacLaren wrote: } } One of the funniest things that I have seen is safety data for water in one } of these books of chemical hazards. It had phrases like: if splashed in } eyes rinse with plenty of WATER, and in case of a spillage wash down drain } with plenty of WATER!!
Ian. here is the copy of that text, sent on this list a few month ago:
Hello all
This is a slightly facetious input re. safety data sheets!
In the UK there is a very useful pair of publications from BDH, a chemicla supply company (used to be known as British Drug House, I believe). These are collcted data sheets. Yes folks, there is one for water. Here are a few salient points which all users of the substance should bear in mind at all times:
1. colourless liquid - maybe that implies it is rare and difficult to see if dropped! 2. Against solubility in water: miscible in all proportions. 3. fire and explosion hazard: not applicable (thats a relief - although if it caught fire I suppose one could foolishly attempt to extinguish with more water?). 4. Health hazard: no significant hazard expected, may be irritating to the eyes. 5. Toxicity: no data. 6. Carcinogenicity: no evidence of carcinogenic properties. 7. First aid - eyes: irrigate thoroughly with water(!) 8. First aid - lungs: remove from exposure. 9. First aid - skin: wash off thoroughly with soap and water (work that one out!). 10. First aid - mouth: wash out thoroughly with water. In severe cases obtain medical attention(!) 11.Reactive hazards: violent reaction with acyl halides, alkali and alkaline earth metals .... 12. Spillage disposal: wear appropriate protective clothing - listed are gloves, goggles, face shield and protective apron. Luckily, respirators are not needed.
We are once again broadcasting Video from the Annual Microscopy & Microanalysis Meeting (this year in Cleveland). Join us virtually on the MSA WWW site
Melanie, I should have mentioned that the 6400F is a field emitter. Thank you.
Jouko, I am sure that charging is not the problem. It even occurred with sputtered gold on carbon resolution samples.
Jan, it sounds as though you were experiencing similar effects. I assume the active field canceling equipment has solved your problem? I would expect the railway to cause the image to shift gradually, peak, and shift back. Not the sudden movement, pause, and sudden return. If related to switching on the motors, I would expect a sudden shift, followed by a gradual return as the train moved off (or the opposite for switching off). JEOL did extensive trouble shooting on my system. They eventually brought in a manufacturer of active field and vibration cancellation equipment as a consultant. My column was placed on an active air table, and inside an active field cancellation system. The conclusion was that the problem was inside the SEM. JEOL eventually exchanged the entire column assembly and the high voltage tank. The difference was like day and night. The resolution and signal to noise were good. The jumping image was gone.
Lately, I have noticed that the jumping image might be returning (hence my inquiry here). I first noticed it at an extraction voltage of 600V (original problem showed up at all voltages). I have seen it at 1kV a couple of times.
JEOL has checked and/or replaced the scintillator a couple of times. The noise canceller has been checked numerous times. The bouncing brightness continues to occur randomly. I have tried flashing the tip when this occurs, and it seems to help. I have not made direct correlation. Perhaps tip instability?
A critical element in imaging the nano-particles is the nature of the support film. Ultra-thin carbon films would be a better bet and you could also try lacey films which would allow you to image the paricles without an underlying substrate.
If you are unable to make these yourself, our company - Ted Pella Inc., is a major manufacturer of support films and we receive many requests for our ultrathin carbon films from researchers studying nano-particles. Please contact us directly if you wish for more information: tedpel-at-aol.com or 1(800) 637-3526.
} I am trying to assist a research group interested in looking at } nanoparticles. Some are made of silver iodide and others are made of } cadmium sulfide, sometimes with a little zinc. They are supposed to be } about 5 micrometers
I got really excited until I saw your correction. I thought that the HVEM would be necessary to see these.
} in size. They do the synthesis and some sorts of } analysis, infrared spectra, I think. They come to my TEM to check on the } size. } } Well, I am having a hard time seeing them.
You could try higher beam current as one responder suggested, but I'd suggest going the other way--use a very sensitive detector and very low beam currents. We have an intensified CCD on our scope which can be used for scanning and focussing with picoamp currents. We also use a small condenser aperture so that no beam is deposited on the sample outside the area being investigated. When the particles have been loca- ted and the image focussed (~1 sec is all that is required), use LoDose film or a slow-scan CCD to record the images. This technique works with TiO2 particles in a similar size range at 1.2 MV (where the contrast is much lower than at 100 kV). Good luck. Yours, Bill Tivol
Mel: You are very correct in letting the microscopy fraternity know about the dangers of dihydrogen monoxide. May I add a further warning that this material which also has another synonym "bis-nor-ethanol" is triphasic and a near universal solvent. Let us be vigilent.
Patrick Echlin
Cambridge, UK also has On Thu, 7 Aug 1997, Melvyn Dickson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues, } } Some eagle-eyed environmentalists have discovered yet another Government } conspiracy to conceal the true harmful nature of a chemical which commonly } contaminates the environment with dire results. } } Full details of this biohazard, dhihydrogen monoxide, can be found at the } following website:- } } http://www.cs.oberlin.edu/students/jbayes/text/dhmo } } If you are properly concerned about the dangers, you may join the coalition } to ban this noxious substance. } } } Mel Dickson } } } } } } } Mel Dickson } Electron Microscope Unit, } University of New South Wales. } Sydney NSW 2052 Australia } } Phone (+612) 9385-2945 } Fax (+612) 9385-1067 } } Website {http://emunit1.babs.unsw.edu.au/emu_top.htm} }
Postdoctoral Research Position in TEM of Semiconductor Epitaxy is Available
Immediately available is a position for a postdoctoral research associate within the Department of Physics at the University of Illinois in Urbana, Illinois, USA. The selected person will study strain distributions in Ge/Si and related epitaxial films by transmission electron microscopy, and will use quantitative measurements of strain to understand the driving forces for island organization and size selection. Both ex-situ microscopy, and in-situ microscopy on our UHV MBE TEM will be involved. The project is supervised by J. Murray Gibson, and involves a collaboration with Jim Coleman, David Cahill and Joe Greene within the UI campus, and the Hewlett Packard Palo Alto Research Laboratory, under NSF funding. The position is for two years (one year at a time, with possible extension to three) and the salary is about $32,000 per year. We are looking for someone with a strong background in conventional diffraction contrast TEM, including the theoretical underpinning and the experimental methods. Experience in semiconductor thin film growth and/or surface science would be an asset. Please communicate by e-mail (preferably) with Murray Gibson at j-gibson-at-uiuc.edu, or call me at 217-333-2997. Note, a second position using our new Low-Energy Electron Microscope and the Scanning Tunneling Microscope on the same project is also available, for which someone with surface science expertize would be most suited.
J. Murray Gibson Professor of Physics and Materials Science Associate Director, Frederick Seitz Materials Research Laboratory University of Illinois, 104 S. Goodwin Ave (Room 258) Urbana, IL 61801 Tel: (217)-333-2997; Fax: (217)-244-2278; j-gibson-at-uiuc.edu
About a year ago I asked for tips on preparation of skin for TEM of stratum corneum. I received some useful replies - Thanks to all!
Now I am looking for references which might show the interaction of medical tapes and dressings with skin, preferably TEM cross sectional images but SEM and LM are interesting as well. So far my literature searches (MEDLINE) have turned up nothing. Any leads would again be appreciated!
Thanks and hope those at MSA are having an enjoyable week!
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
A postdoctoral position at Department of Physics, University of Oslo, is available for a period of two years. The successful candidate will work on a basic research program entitled "Electron microscopy, synchrotron and neutron studies of precipitation and growth in aluminum alloys". The main task will be TEM and/or SANS studies of the early stages of microstructure development in aluminum alloys. Educational requirements include a Ph.D. (or equivalent) in materials science, metallurgy or solid state physics/chemistry and with interest in experimental work. Application deadline is Sept. 15 1997.
For additional information please contact Prof. Tore Amundsen at : tore.amundsen-at-fys.uio.no or Prof. Johan Taft=F8, tel. +47 2285 6113 or +47 2295 8737
A postdoctoral position at Department of Physics, University of Oslo, is available for a period of two years. The successful candidate will work on a basic research program entitled "Electron microscopy, synchrotron and neutron studies of precipitation and growth in aluminum alloys". The main task will be TEM and/or SANS studies of the early stages of microstructure development in aluminum alloys. Educational requirements include a Ph.D. (or equivalent) in materials science, metallurgy or solid state physics/chemistry and with interest in experimental work. Application deadline is Sept. 15 1997.
For additional information please contact Prof. Tore Amundsen at : tore.amundsen-at-fys.uio.no or Prof. Johan Taft=F8, tel. +47 2285 6113 or +47 2295 8737
A postdoctoral position at Department of Physics, University of Oslo, is available for a period of two years. The successful candidate will work on a basic research program entitled "Electron microscopy, synchrotron and neutron studies of precipitation and growth in aluminum alloys". The main task will be TEM and/or SANS studies of the early stages of microstructure development in aluminum alloys. Educational requirements include a Ph.D. (or equivalent) in materials science, metallurgy or solid state physics/chemistry and with interest in experimental work. Application deadline is Sept. 15 1997.
For additional information please contact Prof. Tore Amundsen at : tore.amundsen-at-fys.uio.no or Prof. Johan Taft=F8, tel. +47 2285 6113 or +47 2295 8737
I will be in Cleveland today at the MSA Show. Are there any vendors other than Kevex, BioRad and Evex Analytical that I should visit. I am very eager to see what they have in terms of hardware and software.
Does RuO4 attack glycerol? Also is there a protocol for measuring the concentrations of stained species? I have been working with RuO4 to stain polyvinyl alcohol and need to quantify the concentration / concentration gradient of the polymer. Thanks in advance for any suggestions
Sincerely
Michael Mandanas Particulate Materials Center Pennsylvania State University University Park, PA 16801 mxm67-at-email.psu.edu
Mike, I have only received two responses so far, thanks to both Maria and Lilith, here they are as requested. The first refers to the neurotransmitter GABA.
An additional source I have found for GABAA receptor antibodies is Pharmingen. They carry ABs against alpha 1, 3, 4 and 6. ABs against 3,4 and 6 are suitable only for immunohistochemistry, where anti-alpha 1 is also suitable for western blotting and immunoprecipitation.
On Mon, 4 Aug 1997 16:57:31 -0400 Maria Mejia wrote:
} From: Maria Mejia {maria-at-skivs.ski.org} } Date: Mon, 4 Aug 1997 16:57:31 -0400 } Subject: GABA antibody } To: s.miksys-at-utoronto.ca } }
} Here in our lab. in San Francisco, we are using rabbit } anti-GABA from Sigma cat. # A-2052 with excellent results!! } We did try the GABA from Boehringer Mannheim several times, } but we were never happy with results. Although, we are using } turtle retina tissue - the GABA results are just amazing - } it's a very clean and specific antibody. You have to work } on the right dilution for your application needs, but for us } we use 1:15,000 - overnight -at- RT for 12hrs w/ mild agitation. } We are doing the immuno on frozen sections -at- 10ums thick. } Good luck } maria mejia } smith-kettlewell eye res. inst. } S.F. Ca.
The second response is from Lilith Ohannessian-Barry from the National Research Council Institute of Biological Sciences in Ottawa. The NRC has a very good antibody to alpha 1 subunit, but they have no vendor as yet. If any-one is interested I can put you in touch with Lilith.
I am trying to heat a glass powder which contains an organic binder to= =20 1000 degrees C to observe the binder burnout and glass transition=20 temperatures. However, I need advice on how to prevent the powder=20 from moving during heating. The particles eventually move around,=20 which wouldn't be such a problem except that they often "jump" up onto= =20 the heat shield and obliterate the field of view. I hesitate to=20 adhere the particles to the alumina crucible, since anything I use=20 might react with the binder which is already present in the powder (I=20 do not know what the binder is). =20 Any suggestions would be greatly appreciated. =20 TIA, =20 Leslie Link e-mail: Leslie.Link-at-us.gtc.boc.com
I have been looking (by TEM) at an alloy with mostly Ni, Al,Pt. The alloy was cycled to high and low temperatures several times. The resulting microstructure is highly twinned and it resembles the microstructures seen in martensitic Fe alloys. We have not been able to find other references which would indicate that Ni, Al,Pt alloys undergo martensitic transformations. I would appreciate any suggestions or references that might be useful.
Does anyone have any old Latico "T" series condensor lenses for our Durst Enlarger (SM183) they would be willing to give up? We would pay fair price for any or all of 240T, 200T, 130T and 85T. Thanks!
I would like to announce that the URL of my series of WWW pages called "Microscopy and Imaging Resources on the WWW" has been changed. The University of Arizona College of Pharmacy (which hosts these pages) has decided to retire their old domain name and move to a new one. Currently all page requests for a URL using the old name of "www.pharm.arizona.edu" are being forwarded automatically to "www.pharmacy.arizona.edu", BUT this forwarding will cease in about 6 months.
In preparation for this announcement I have updated the pages to clean out dead links and add new ones I've located. I may eventually stoop to using the dreaded {BLINK} tag near the top of each page to remind visitors to check their links (I promise I'll wait until the end is near).
I appreciate those of you who are the sources of many of the links on my pages. My goal for these pages has been to "collect relevant sources that provide educational and technical information about biological microscopy and imaging in a manner that is non-commercial (in the case of information from vendors) and clear enough for graduate students who are neophytes to microscopy and imaging." If you know of other resources that I'm missing, please contact me.
If you have a reciprocal link to one of my pages I will try to contact you directly to remind you of this change.
Thanks to all who are regular visitors or who have linked to these pages. This project is an outgrowth of my position as manager of the Experimental Pathology Service Core for the National Institutes of Environmental Health Sciences (NIEHS) funded Southwest Environmental Health Sciences Center (SWEHSC). NIEHS is very strong on community outreach and education and I figure these WWW meta-lists fit in rather nicely with that goal.
THE CORRECT URLs ARE:
Microscopy and Imaging Resources on the WWW http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/m-i_onw3. html
Histology on the WWW http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/w3-histo. html
Use & Misuse of Formaldehyde Fixatives
http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/formalde. html
Other Commonly used Histologic Fixatives
http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/otherfix. html
Confocal on the WWW http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/conf_www. html
Electron Microscopy on the WWW http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/em_w3.html
Digital Imaging on the WWW http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/dig_imag. html
Free Publications of Interest to Microscopists http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/freemags. html
Reciprocal Links http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/m-i_link. html
Reference Material for Biological Scientists http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/referenc. html
(There are a few other pages, but they are mostly of interest to local folks.)
Yours, Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html
Dear Jordi, Seeing something resembling martensite under TEM does not necessarily mean the alloy has undergone martensitic transformation. Actually, it is quite common to see the twinned structure which may not be a result of martensitic transformation in many high temperature alloys such as Ti-Al, Ti-Al-Nb, Ni-Al and so on. Twins may form through phase transformations including martensitic transformation, or mechanically/thermally introduced strains. Your case seems to be most explanable by the later.
There are two most important features of martensitic transformation which I can remember by now. One is the speediness of transformation and the other is a certain crystallographic relationship between parent phase and newly formed martensite.
Chao-Ying Ni Microscopy Division Batta Laboratories, Inc. Newark, DE 19713
On Wed, 13 Aug 1997, Marti, Jordi wrote: } } I have been looking (by TEM) at an alloy with mostly Ni, Al,Pt. The alloy } was cycled to high and low temperatures several times. The resulting } microstructure is highly twinned and it resembles the microstructures seen } in martensitic Fe alloys. We have not been able to find other references } which would indicate that Ni, Al,Pt alloys undergo martensitic } transformations. I would appreciate any suggestions or references that } might be useful. } } Thank you, } } Jordi Marti }
Does anyone have any experience with the SEM/EDS analysis of soot resulting from incomplete combustion? Can differences be detected in soot that results from different fuel sources such as wood, oil burning furnaces, natural gas, LP gas etc.
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Does anyone have any experience with the SEM/EDS analysis of soot resulting from incomplete combustion? Can differences be detected in soot that results from different fuel sources such as wood, oil burning furnaces, natural gas, LP gas etc.
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All,
We are getting ready to order the Kevex upgrade to replace our existing IOMEGA 10 Meg drives with the dual Syquest 44 Meg drives on our 9 year old Kevex system.
I was wondering if anyone else had done this and if there were any tips, tricks or warnings with installation or use of these drives?
Thanks for any words of wisdom!!
John Giles Senior Materials Engineer Honeywell Space Systems
Does RuO4 attack glycerol? Also is there a protocol for measuring the concentrations of stained species? I have been working with RuO4 to stain polyvinyl alcohol and need to quantify the concentration / concentration gradient of the polymer. Thanks in advance for any suggestions
Sincerely
Michael Mandanas Particulate Materials Center Pennsylvania State University University Park, PA 16801 mxm67-at-email.psu.edu
Jake, I also have bright horizontal streaks flash across the screen, sometimes. These are caused by emission noise, and a "flash" usually clears them up (unless they are from a charging sample). The problem I was referring to, shows up as "bands" of different levels of brightness (i.e. Start a slow scan; maybe the first 1/2" is one brightness level, the next 1/4" is brighter by several shades of gray, the next 3/4" may be back to the original level of brightness. The widths, and positions, of the bands are random, I only tried to illustrate the problem). The sudden lateral movements of the image are while remaining in super rapid. The shifts when switching from super rapid to slow, and back, are due to changing blocks of circuitry in the console. These shifts can be minimized by your service people.
I've received several private comments about the URLs I posted yesterday. Many have said that the links didn't seem to work. I suspect that in most cases the rather long URLs have "wrapped" from one line to the next. I think you'll find that if you include the part of the URL text that wrapped onto the next line that the links will work (all of them work for me). I'm sorry the URLs are so long, unfortunately the same folks that changed the server name on me won't let me have shorter URLs.
Yours, Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html
They kept trying to say that my problems were fields, but the active field cancellation could not affect the visual symptoms. The ultimate proof that it was inside the system, is the fact that the problems went away when they replaced the entire column. It is easy for SEM manufacturers to blame fields for problems they can not trace, or easily solve. They all seem to use this crutch from time to time.
Do not get me wrong, I know fields can be a problem. My response to that is that, as the systems become more and more susceptible to the ocean of magnetic noise they live in, the manufacturers need to prevent the affects with their design, and not use it as an excuse for poor performance in the real world.
Enough of that! I was just wondering if anyone else was having these problems on similar systems. It seems as though there are a few out there.
Greetings to all, I do TEM on human tissues ... mostly renal biopsies. I have noticed that the past 2 or 3 that I have processed have had tiny "pin holes" when viewed under the scope ... don't know why! I have changed NOTHING in my procedure, but feel this is a processing problem. (I hand process everything.) I use Spurr's for embedding ... could one of the four ingredients have gotten contaminated somehow ... maybe with moisture? Should I open all new bottles and try that? Maybe air bubbles are formed when I mix/stir the Spurr's ... that's never happened before when I have mixed it. Has anyone else had this experience just happen "out of the blue" like this? The "pin holes" are so tiny that they don't even show up in cutting the thin sections at 70nm ... only after being stained and put on the 'scope at the END of the whole thing! This doesn't hamper the diagnosis at all, but it's not the quality that I'm used to giving our pathologist and I'm not a bit happy about it! What do you think? Thanks in advance for your help.
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
I am writing from the St. Louis Science Center in St. Louis, MO. The Science Center is a non-profit, hands-on science museum that is free to the public. We have a gallery that is opening next month and we need some good EM and/or light microscopy photographs that show various examples of cell morphology. We would like both animal and plant cell photographs. Needless to say, the photos will have to be of excellent quality, because they will be installed in the gallery and will be on display for the life of the gallery (a minimum of five years). We will be displaying the photographs blown up to at least 8X12 inches as back-lit images on a kiosk.
Does anyone out there have anything to share with us? Please e-mail me directly at cencarna-at-slsc.org. Thank you so much for your help.
Cindy H. Encarnaci=F3n, Ph.D. St. Louis Science Center
A post - doctoral research associate position exists within the interface science group of the Chemistry and Materials Science Directorate of Lawrence Livermore National Laboratory. The group studies in a fundamental research program the relation between the structure and composition of internal interfaces and their properties, including mechanical, diffusional, and mobility. The materials of interest are primarily metals, but interfaces in ceramics and metal/ceramic heterophase interfaces are also studied. The position is for an experimentalist to characterize and quantify grain boundaries and interfaces in regards to their structure, morphology, segregation of minor species, and diffusion. Model interfaces are fabricated for study by diffusion bonding oriented single crystals in our ultra-high vacuum diffusion bonding machine. A primary tool for characterization will be the transmission electron microscope (TEM), but it is anticipated that many other methods could be brought to bear on issues raised by this work. However, conventional and high resolution imaging and analytical techniques will be pushed to their limits by this research. Quantification in the data analysis will require computer code development.
The applicant will be expected to hold a PhD in materials science or a closely related field. A thorough understanding of crystal defects is essential. The applicant will be expected to have extensive experience with TEM. Experience with computer code development, IDL, FORTRAN, and the UNIX operating system is highly desirable. Excellent oral and written English skills are essential. The initial appointment is for one year, with extensions possible for one or more additional years.
Those interested, please contact:
Dr. Geoffrey H. Campbell Lawrence Livermore National Lab Mailstop L-356 P.O. Box 808 Livermore, CA 94551-9900 USA Phone:(510) 423 - 8276 FAX:(510) 424 - 4737 e-mail:ghcampbell-at-llnl.gov
Yes we have had similar "out of the blue" problems with Spurrs on several occasions.
The DMAE component of Spurrs may well have gone bad, it seems to be the most suseptible. I write the date all the bottles when I open them now, in case I am in any doubt. Also check the caps are still good, as I find they occassionally tend to split open in storage.
Another problem we had probably wouldn't apply, as it was caused by a failing seal on our automatic tissue processor: an increase in prop. oxide evaporation during processing resulted in incomplete infiltration.
Hope this helps!
Miss A.J.Wilson Electron Microscope Unit St George's Hospital Medical School Cranmer Terrace Tooting London SW17 ORE Tel: 0181 725 5220 awilson-at-sghms.ac.uk awilson-at-aw.u-net.com
It has been suggested that I should copy this message to all subscribers.
------- Forwarded Message Follows -------
I recently upgraded my Kevex 8000 to use the Syquest drives. Everything went smoothly and the new drive system is working great. The kevex upgrade struck me as somewhat pricy, considering the actual "street price" of the drives, but insures you get the proper cables, support, setup, etc.... One disadvantage is that the 230Mb (for PCs) drive becomes a 44 Mb, DEC/RT11 formatted system.
Eventually, I expect to stop using my 44Mb Bernoullis entirely. To ensure easy availability of recent data during a transition time, my system is currently configured to run one SyQuest drive and the (old) dual 44Mb Bernoullis. Hardware limitations prevented me from running both SyQuests and the pair of 44s.
FWIW... My office PC was setup with an internal 44Ber and an Adaptec 1542 so that I can read the RT11 format data into a DOS/Win system. Using the "spare" SyQuest, I determined that the PC system will support both at the same time.
Woody White Mcdermott Technology, Inc http://www.mtiresearch.com/
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All,
We are getting ready to order the Kevex upgrade to replace our existing IOMEGA 10 Meg drives with the dual Syquest 44 Meg drives on our 9 year old Kevex system.
I was wondering if anyone else had done this and if there were any tips, tricks or warnings with installation or use of these drives?
Thanks for any words of wisdom!!
John Giles Senior Materials Engineer Honeywell Space Systems
Hi: Since many of the subscribers of the Microscopy mailing list have been asking me about the First Electronic Conference in Analytical Chemistry I have decided to post the following message to explain a little more about it...
************************************** Analytical Chemistry Mailing List ************************************** Due to the success of previous virtual conferences (MGM EC1 & EC2, first and second electronic molecular graphics and modelling conference http://bellatrix.pcl.ox.ac.uk/egc/ and http://bellatrix.pcl.ox.ac.uk/egc2/home.html, ECHET96, electronic conference on heterocyclic chemistry, http://www.ch.ic.ac.uk/ectoc/ehet96/, ECCC1 & ECCC2, first and second electronic computational chemistry conference http://hackberry.chem.niu.edu:70/0/ECCC/homepage.html) it was decided to set up a moderated mailing list to pave the way for the
FIRST ELECTRONIC ANALYTICAL CHEMISTRY CONFERENCE to be held in November of this year (3rd-14th)
Preperation for the electronic conference is still in its early stages and any input (discussion topics, suggested conveners, short courses literature/product reviews etc.) would be greatly appreciated.
You can add your name to this mailing list by sending an e-mail to
ac-request-at-vei.co.uk
leaving the subject line blank and
subscribe {your_email_address}
as the message body (no signatures). Once accepted you will be sent a mail welcoming you to the mailing list and explaining how to post a message.
Among some of the areas we hope to include in discussion will be topical aspects of: * NMR, IR, UV, HPLC and GC. * Electrochemistry. * AA, ICP and ICPMS. * Macromolecular and Small molecule Crystallography. * Analytical techniques. * Method development and validation procedures. * Bioanalytical forum. * MS and applied topics. * Capillary Chromatography & Electrophoresis. * Statistical analysis of data. * Microscopy
Any suggested additions may be made through the mailing list above.
Authors can opt to have their presentation in the following categories:
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During the conference interaction, presentations and discussions will take place via the Internet using a Java-based virtual conference centre, WWW-based discussion forums and an electronic mailing list. Before the conference, a timetable for lectures and discussion sessions for each section will be posted. Since these realtime discussions are an integral part of the conference, authors will be expected to attend one for their subject; the right is reserved not to referee submissions by authors who do not attend one of these sessions.
The Conference will feature a Virtual Exhibition where exhibitors will be able to describe the activities of their organization, display their products and services and interact with registrants. Potential exhibitors should contact the conference organiser.
Here are some basic suggestions for this king of problem: It could be water in you solutions - try new chemicals, but don't throw the old ones away until you're sure they were the problem. OR it could be a change in the size of the tissue? or the fixation process, ie the fix is old. Maybe try putting the Spurrs under a vacuum before you use it, or put the tissue and Spurrs under a SLIGHT vacuum before you embed it. Do you use propylene oxide to dehydrate? I have had some old prop. go bad on me and make all the blocks soft. If it had a small amount of water in it, it might cause your problem. Good luck.
Karen Pawlowski Sr. Research Assoc. ENT Res. Lab., UT Southwestern Medical Center PhD student, UT Dallas Dallas, TX
On Thu, 14 Aug 1997, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings to all, } I do TEM on human tissues ... mostly renal biopsies. I have noticed } that the past 2 or 3 that I have processed have had tiny "pin holes" } when viewed under the scope ... don't know why! I have changed NOTHING } in my procedure, but feel this is a processing problem. (I hand process } everything.) I use Spurr's for embedding ... could one of the four } ingredients have gotten contaminated somehow ... maybe with moisture? } Should I open all new bottles and try that? Maybe air bubbles are } formed when I mix/stir the Spurr's ... that's never happened before when } I have mixed it. Has anyone else had this experience just happen "out } of the blue" like this? The "pin holes" are so tiny that they don't } even show up in cutting the thin sections at 70nm ... only after being } stained and put on the 'scope at the END of the whole thing! This } doesn't hamper the diagnosis at all, but it's not the quality that I'm } used to giving our pathologist and I'm not a bit happy about it! What } do you think? } Thanks in advance for your help. } } Sharron G. Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas }
If Spurr's resin has served you well in the past, I stick (sorry) with it. I'd recommend you get a fresh kit from any of the suppliers out there who handle the stuff. In our lab, where the humidity is condensing from May until snowfall, we have at least one bout of bad resin nearly every summer. My own suspect is that the anhydride winds up not so after a few people leave the lid loose, either on the stock bottle, or on an individual preparation. We also refrigerate our mixtures to extend the pot life and despite my warnings I'll bet there are those who don't let them reach the dew point (i.e. RT) before opening. Given the relatively low price of the kits, I'd reckon it's worth one's time to get a fresh one.
With respect to mixing resins, Spurr's is an epoxy mix to begin with, but if you don't like the characteristics of the published formulas, you can cook up your own. I've taken to using Quetol 651 in an equiequivalent mix with the VCD instead of the DER resin. We call it Spurtol. It seems to stain a little easier than the DER softened mix. DMAE works as an accelerator just as in A.Spurr's original. Works well with plants, chicken, and some high density polymers.
cheers, John Heckman TEM Supervisor/ Academic Specialist MSU Center for Electron Optics Michigan State University
Hi Folks, A little good news for those of us unwilling/unable to pay $6 billion for a RIP to network the Fujix printer. A company called Techpool software in Cleveland (www.techpool.com) have produced a RIP which costs $490 and seems to work pretty darn well. This allows printing from any application or across the network. The way the networked system works is like the Lasergraphics sildemaker, you mount a "hot" or watched folder and save printjobs to that folder and a TSR on the computer with the printer watches that queue for jobs. Works pretty well in our hands (though we are still using the demo, which you can get from the web site)
BTW I have no interest in this company etc etc Simon
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
Do any of you have experience labeling cell surface antigens for SEM? I've been working with someone who has a couple of these projects in mind. We made one attempt at gold labeling in the standard fashion but were thwarted by the low resolution of my non vibration-mounted scope. Or maybe the labeling didn't work in the first place.
I think this person will have to go elsewhere to get what she needs because of the limitations of my scope but I told her I'd post a request for established protocols and any tips you might have to offer. The particular experiment she has in mind is to visualize annexin 4 on the surface of HUVEC cells.
Thanks in advance for your help, Tori
Tori Hatch thatch-at-hsph.harvard.edu Physiology Program Harvard School of Public Health 665 Huntington Ave. Boston MA 02115
Just so that every one knows Taab 812 is ~6 times more expensive than the conventional spurr or epon... Neelima Shah
At 12:58 PM 8/15/97 GMT+0200, ROBIN CROSS wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The following is posted as a favor to one of the companies which represents our products.
Pulcir, Inc. has an opening for a sales/support engineer in the Southeastern U.S. with experience in electron microscopy and EDS. If interested please contact:
A six month maternity-cover postdoc. position is available in my group from 1.10.97. Applicants require some knowledge of cell culture and electron microscopy to investigate the interaction between living tissues and new glass-ceramic biomaterials with dental and orthopaedic uses.
Further information and application details are available from:
The Director of Human Resource Management University of Sheffield Western Bank SHEFFIELD S10 2TN email: Jobs-at-sheffield.ac.uk
Quoting refererence R1286
I am happy to deal with informal questions.
Dr Paul V. Hatton Lecturer in Biomaterials School of Clinical Dentistry University of Sheffield Claremont Crescent SHEFFIELD S10 2TA
Tel. (0114) 271 7938 Fax. (0114) 2665326 or 2797050
It has been pointed out to me, and rightfully so, in my previous posting about the price of Taab as compared to Epon or Spurr I neglected to state that the cost in USA. I have no personal or financial interest any one of the products but since in recent times cost has become a major concern in most institutions i thought it necessay to point out. I apologize for my haste in posting. Neelima Shah Regards... :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ;-) ;-) Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.htm
MSA is collaborating with the Lawrence Hall of Science, an outstainding science education center, to publish a classroom manual as part of Project MICRO, MSA's middle school educational outreach program. It's titled "Microscopic Explorations", and publication is scheduled for May '98. LHS materials are used in 25% of U.S. schools, so it will get wide distribution. The LHS has asked for color micrographs to use on all four covers (inside & outside, front & back). We need LM more than EM, at fairly low magnifications, and we need them by SEPTEMBER 25. Topics presented in the manual are preferred: pond life, brine shrimp, insects, crystals of common substances, sand grains, fabrics, color printing - but other eyecatching things that are familiar to children are welcome. Your contribution will be acknowledged in the manual. Please let me know directly, ASAP, if you are interested in sending an image.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
this is not my field but I do know of someone who had problems with gold labelling on the SEM.
You haven't given much information: 1.The name of your microscope and resolution 2. The size of gold particle (eg if you are using 5nm gold they are extremely difficult to see against a thick SEM sample not just because of resolution but because of insufficient thickness to scatter sufficient electrons). If you must use small gold I believe that silver enhancement might be possible. 3. I assume you have only used carbon to coat the samples because most metal coatings will make the gold invisible or difficult to see. 4. Do you have a backscattered detector? Because they are much better at picking up the mass difference of the gold against the lighter elements of the specimen. 5. You could check to see if you can detect the gold on its own by drying some onto the stub - this would at least tell you if it's a resolution problem. If this works you could try to find a positive control and look for the label on that.
If you have done all of the above then I would wait for a real expert to answer.
I hope this helps - but as I said I have never used gold labelling on the SEM.
Malcolm Haswell e.m. unit University of Sunderland UK
Does RuO4 attack glycerol? Also is there a protocol for measuring the concentrations of stained species? I have been working with RuO4 to stain polyvinyl alcohol and need to quantify the concentration / concentration gradient of the polymer. Thanks in advance for any suggestions
Sincerely
Michael Mandanas Particulate Materials Center Pennsylvania State University University Park, PA 16801 mxm67-at-email.psu.edu
For those of you who read the American Lab coverage on Microscopy & Microanalysis '97 and were confused about the source of the book "Optimizing Light Microscopy for Biological and Clinical Laboratories": The book is not available through MSA but is available through MME (Microscopy/Microscopy Education). If you are interested in ordering, please send your Fax number to: Barbara Foster MME (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com We will fax an order form to you, and will honor the 20% M&M '97 discount through Labor Day.
Our apologies for the editorial mix up in both email and annotation.
Barbara Foster Consulting editor to American Lab Consortium President, Microscopy/Microscopy Education
A quick "THANKS!!!" to the nearly 30 companies who contributed to the article covering the Microscopy & Microanalysis '97 meeting. American Lab gave us a whopping 8 page coverage! Several attendees stopped by the MME booth to say that they never knew the meeting existed before the article appeared and several members of MSA council expressed their great appreciation at having such wonderful promotion. For those of you who have not yet seen the article, look for American Laboratory, July, 1997, pp. 38-46.
A reminder that this article was actually part of an on-going column called "Focus on Microscopy" which appears approximately bimonthly in Am. Lab. Contributions are always welcome. Our next column will be on managing images on both Intra and Internet. Please send suggestions and helpful hints to: Barbara Foster Microscopy/Marketing & Education 53 Eton Street Springfield, MA 01108 Ph: (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com
Thanks again! Barbara Foster Consortium President, MME Consulting editor, American Lab
You should first ask how the larger disks will be handled. RT-11 and TSX+ were limited to volume sizes of 32 MB under the DU driver. There was an enhanced DU driver produced by a third party that would handle larger drives as a single volume.
When we added a hard drive to our Delta, we limited it to 170 MB because we were keeping 2 Bernoulli drives which would leave only 6 of the 8 possible DU units of up to 32 MB each to use for the hard drive. Thus anything more than 192 MB would have been wasted space.
There may also be a controller issue. The original Bernoullis on our Delta used a SCSI controller that required the DL driver. That driver supported up to four 10 MB devices. Your new disks will probably require a switch to a DU-type controller (see above) like we had to purchase for our upgrade. Such cards should be available on the used market now at a decent price.
Feel free to ask for more details.
At 08:33 AM 8/14/97 -0500, you wrote: } All, } } We are getting ready to order the Kevex upgrade to replace our existing IOMEGA } 10 Meg drives with the dual Syquest 44 Meg drives on our 9 year old Kevex } system. } } I was wondering if anyone else had done this and if there were any tips, tricks } or warnings with installation or use of these drives? } } Thanks for any words of wisdom!! } } John Giles } Senior Materials Engineer } Honeywell Space Systems ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
NIH Image for Macintosh may work for your application. It does have video image capture capability. This is a public domain piece of software. The following is an excerpt from an e-mail about 3-d reconstruction and has the URL from which to download the MAC and Win95 versions of this software.
NIH-IMAGE This popular freeware program for image analyses is originally written for the Mac, but now is also available as Win95 program. Download: http://www.zippy.nimh.nih.gov/ (Mac) or http://www.scioncorp.com/ (Win95) Many information e.g. online manual, macros, example-files and additional download possibilities can be found at: http://rsb.info.nih.gov/nih-image/ As an example animated reconstructions of plant cells (based on semithin
sections) can be found on the home page of Gary Chinga: http://www.nvg.unit.no/~gary Information about the NIH-Image mailing list can be found at http://www.soils.agri.umn.edu/infoserv/lists/nih-image/
Good Luck
============================ Michael D. Standing Electron Microscopy Technologist Brigham Young University ============================
mauty-at-DAIRY.TEAGASC.IE wrote:
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } } I have a Matrox Meteor imaging board and would like to acquire RGB } images via a JVC TK1070E CCD camera fitted to a Zeiss confocal } microscope. Does anyone know of any shareware (or fairly cheap) image } acquisition software that I could use to view and acquire images, } preferably as TIFF image ? } } Regards } } Mark Auty } DPC Moorepark } Fermoy } Co. Cork, Ireland } mauty-at-dpc.teagasc.ie
Using the programs named RTDIR.EXE and RTCOPY.EXE (which were a portion of the old Kevex "Report Manager" software package) I can read a 44 Mb Bernoulli or a SyQuest (RT format) disk into the PC clone. The two programs operate much like DOS's DIR and COPY commands. An Adaptec 1542 SCSI controller is used to interface the two drives to the PB bus.
Given this... I can transfer files ok, but the EDS spectra are packed in "library" files and I haven't tried to disect one. It has been a while, but I think you can SAVE/EXTERNAL a spectrum which would (like maps,images etc.) be a discrete file.
Woody White Mcdermott Technology, Inc.
-------------------------------------
Woody,
I appreciate the reply, and have a question for you about your office PC. It sounds like you are able to import the Kevex data into a PC and make use of it. My technician was very interested in your reply and wondered what you are importing with the RT11 data. Specifically, are you able to read the RT-11 data from an individual spectrum on your PC? We have a PC linked to our Kevex system to import Feature analysis data, but it would be nice to be able to import the spectra to the PC also so you could use in a report.
I recently upgraded my Kevex 8000 to use the Syquest drives. Everything went smoothly and the new drive system is working great. The kevex upgrade struck me as somewhat pricy, considering the actual "street price" of the drives, but insures you get the proper cables, support, setup, etc.... One disadvantage is that the 230Mb (for PCs) drive becomes a 44 Mb, DEC/RT11 formatted system.
Eventually, I expect to stop using my 44Mb Bernoullis entirely. To ensure easy availability of recent data during a transition time, my system is currently configured to run one SyQuest drive and the (old) dual 44Mb Bernoullis. Hardware limitations prevented me from running both SyQuests and the pair of 44s.
FWIW... My office PC was setup with an internal 44Ber and an Adaptec 1542 so that I can read the RT11 format data into a DOS/Win system. Using the "spare" SyQuest, I determined that the PC system will support both at the same time.
Woody White Mcdermott Technology, Inc http://www.mtiresearch.com/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
All,
We are getting ready to order the Kevex upgrade to replace our existing IOMEGA 10 Meg drives with the dual Syquest 44 Meg drives on our 9 year old Kevex system.
I was wondering if anyone else had done this and if there were any tips, tricks or warnings with installation or use of these drives?
Thanks for any words of wisdom!!
John Giles Senior Materials Engineer Honeywell Space Systems
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mark_munro-at-bio-rad.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Does anyone know of any good sources of information on the emission } and absorption spectra of different fluorochromes, and their binding } properties? } } Thanks in advance, } } Mark Munro.Mark,
An excellent reference is the "Handbook of Fluorescent Probes and Research Chemicals" First copy usually free; subsquent ones, nominal cost. Contact: Molecular Probes, Inc. P O Box 22010 Eugene, OR 97402 Ph: (503)465-8353 Fax: (503)344-6504
Best of luck... and if you need further info on fluorescence, please give us a call or email.
Barbara Foster Consortium President Microscopy/Marketing & Education 53 Eton Street Springfield, MA 01108 Ph: (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com
Thank you to all who responded to my TEM artifact problem regarding "pinholes" in the specimen. The suggestions were very helpful and I think the culprit is the bottle of absolute ETOH I have been using. I opened a new bottle today and am processing a renal case. If this doesn't do it, I'll go to the next suggestion and use new Spurr's components. (Someone asked if the biopsies were put in those blue sponges that also cause holes in tissue ... no, the biopsies I get are put straight into small bottles of fixative ... without sponges or cassettes.) Hopefully, the new bottle of ETOH will solve the problem. Thanks, again for your thoughts ... Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
Mark - try Molecular Probes, Inc. http://www.probes.com (514)456-8353 they know about fluorochromes! -Mike
On Mon, 18 Aug 1997 mark_munro-at-bio-rad.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Does anyone know of any good sources of information on the emission } and absorption spectra of different fluorochromes, and their binding } properties? } } Thanks in advance, } } Mark Munro. } } }
The following is posted for your consideration. Bob Craig OSRAM SYLVANIA Products Inc.
Scanning Electron Microscopy/X-ray Microanalysis Specialist
Exceptional opportunity exists for a qualified individual to apply her/his analytical and experimental skills in support of research, development, and manufacture of sophisticated lighting products.
This position is responsible for the application of scanning electron microscopy and X-ray microanalysis to study materials problems associated with the development and manufacture of state-of-the-art lighting products. Responsibilities will include: developing new methods for evaluating lamp materials, performing in-depth studies to solve complex lamp materials problems, and operating and maintaining associated analytical instrumentation. The successful candidate will be expected to function as part of a highly skilled technical team, interact in a consultatory fashion with internal R&D and manufacturing customers, report results and make technical recommendations based on the interpretation of those results.
Candidates with the following educational background will be considered: a M.S. degree (research) with 3-5 years experience or a recent Ph.D. in chemistry, materials science or physics. Hands on experience in scanning electron microscopy and X-ray microanalysis of inorganic materials, particularly metals and ceramics, is an absolute must. In addition, the successful candidate should have a firm knowledge of electron-solid interactions and a thorough understanding of the physics of X-ray generation and interaction with crystalline solids. Experience in X-ray diffraction and thermal analytical techniques is also desirable. Good inter-personal, and writing skills are requisite.
Please respond with a resum=E9 to: Mr. Amando Llorente, Human Resource Manager OSRAM SYLVANIA Products Inc. Lighting Research Center 71 Cherry Hill Drive Beverly, MA 01915
I was asked to post the following position announcement by a collegue. The institution would like to fill it for the upcoming academic year.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Full-time Assistant Professor of Biology Academic Year 1997-98
Teaching responsibilities to include:
Introductory Biology
and
Human Anatomy & Physiology
and/or
Integrated Physical/ Biological Science
and/or
Introductory Electron Microscopy
and/or
Histology
Teaching load approximately 12 credit hours per semester. Master's degree required; Ph.D. preferred. New position; may be renewed pending funding. Send letter of interest, curriculum vitae, transcripts and list of references to:
John F. May, Ph.D., Coordinator Department of Biology Marian College 45 South National Ave. Fond du Lac, WI 54935
To those of you trying to respond to our earlier message of thanks for the American Lab article: We are having trouble with our email. Please do not use the "RE:" method of responding. Email directly to: mme-at-map.com.
To those of you trying to get through to Microscopy/Microscopy Education about this book: We are having email problems. Please do not respond using the "RE" function. Send email to: mme-at-map.com.
The techniques listed on the board almost to a person refer to dehydrating the specimen after removing the wax with xylene and then processing as usual. It should be noted that several of my colleagues as well as myself have processed the tissue without rehydrating and then dehydrating the tissue again. The key question is WHAT are you possibly gaining b;y rehydrating the tissue????? The only possible reason or certainly the main one would be to try and "osmicate" the tissue. It has been our observation that the tissue simply does not turn black or pick up any significant amoun of osmium whatsoever after this rehydration. I think that this "old technique" to recover paraffinized tissue is based as much on "urban legend" as anything. The least amount of processing the better. Uranyl Acetate is soluble in absolute acetone for example although en bloc staining of tissue here seems mostly futile. Better to process the tissue directly into plastic after xylene by washing in acetone and propylene oxide and infiltrating into plastic. Afterwords use heavy duty uranium and lead stain ( also I guess one could expose the grids to Osmium vapors to attempt to "osmicate" but I suspect the effort might not be worth it. I would apprecitate some serious comment on this including wheter you have tried this more direct approach. May you never have to do this as I and others have had to for the last 15 years. bob
I am trying to measure the thickness of corneas. Currently, I am using a pachymeter(SD=10 microns). The method is not accurate because it imply compression of the tissue. Could I expect a lower standard deviation from measurements obtained from x-sections using LM? Is anyone familiar with a method that would not involve LM (tissues would not be processed for LM a suitable alternative is employed)? I read a paper where the authors used the universal measuring microscope on corneas embedded in epon. What does that mean? What is is? Could it be good for my purpose?
Thanks in advance,
Dan Caruso Biological Technician medjet-at-worldnet.att
Mark, go into the links on our site. Use search to find 'fluoro' (there are over 400 links) and this will take you to a link at the uni of Buffalo with an extensive table. Just what you are looking for. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au ____________________________________ } Does anyone know of any good sources of information on the emission } and absorption spectra of different fluorochromes, and their binding
} properties? } } Thanks in advance, } } Mark Munro. } }
} } It has been pointed out to me, and rightfully so, in my previous posting } about the price of Taab as compared to Epon or Spurr I neglected to state } that the cost in USA. I have no personal or financial interest any one of } the products but since in recent times cost has become a major concern in } most institutions i thought it necessay to point out. I apologize for my } haste in posting. } Neelima Shah } Regards... } :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ;-) ;-) } Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.htm } } Regards... :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ;-) ;-) Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.htm
Friends, We have been notified that our histology service lab here at the University of Arizona may be audited. Because I "inherited" the pricing structure from someone else, the documentation to justify our prices is somewhat lacking. I'm working on that now, but in the meantime would someone be willing to compare notes with me that's gone through such a process?
We would particularly be interested in corresponding with a lab that serves a similar constituency to ours. We serve only University affiliated researchers (we're in the medical college). We do a wide variety of research specimens (mostly the usual rodent tissues, but occasional insects & botanicals), but no clinical work. For more on what we, do our Web site is: http://www.cba.arizona.edu/histology-lab.html
If you can assist me or refer me to someone who can assist me I'd greatly appreciate it. Since this issue may not be of interest to most of the subscribers to this list, please reply to me privately. Thank you.
Yours, Doug Cromey Supervisor, Cell Biology & Anatomy Histology Service Core Lab ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html
Friends, We have been notified that our histology service lab here at the University of Arizona may be audited. Because I "inherited" the pricing structure from someone else, the documentation to justify our prices is somewhat lacking. I'm working on that now, but in the meantime would someone be willing to compare notes with me that's gone through such a process?
We would particularly be interested in corresponding with a lab that serves a similar constituency to ours. We serve only University affiliated researchers (we're in the medical college). We do a wide variety of research specimens (mostly the usual rodent tissues, but occasional insects & botanicals), but no clinical work. For more on what we, do our Web site is: http://www.cba.arizona.edu/histology-lab.html
If you can assist me or refer me to someone who can assist me I'd greatly appreciate it. Since this issue may not be of interest to most of the subscribers to this list, please reply to me privately. Thank you.
Yours, Doug Cromey Supervisor, Cell Biology & Anatomy Histology Service Core Lab ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html
Digital Instruments (DI) will be hosting two workshops on Scanning Probe Microscopy in the New England area during the month of September, 1997. The workshops will consist of two lectures in the morning, and a hands-on demonstration in the afternoon. Samples of interest to attendees may be analysed at this time. Those interested in attending these free workshops can get more information at the DI web site (www.di.com) or can contact Rich Goodheart at 410-437-1805.
Eric wrote: ========================================== To the wealth of knowledge out there on the list.. Does anyone know if the Zerostat guns are still available anywhere?? ========================================== They ARE still available, not just from SPI but also from several of our friendly competitors. Remember, Zerostat (R) is a registered trade name and should be indicated as such.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The Zerostat guns have been discontinued for a few years now however we have now reintroduced them into our line. Please contact us for further information. Stacie Kirsch Electron Microscopy Sciences Tel: 215-646-1566
A full-time microscopy technician position is available in the laboratory of Dr. Ann Hubbard at The Johns Hopkins School of Medicine. The position involves biological specimen preparation, imaging, and image analysis at the light and electron microscopic level. The research environment in the schools of Medicine and Arts and Sciences at Hopkins encourages a cooperative approach that is reflected in collaborations among diverse labs, faculty, students, post-doctoral fellows, and technicians. Thus, there is a potential for interactions with technical and academic personnel throughout the institution.
Qualifications
The successful applicant will have a BS/BA in a biological science and at least 2 years of experience in a wide variety of microscopy techniques. Proficiency in fixation and embedding, ultramicrotomy, immunocytochemistry, and digital and photographic imaging techniques is essential. Experience in the operation of TEMs and the operation and maintenance of research-grade light microscopes is required. Preference will be given to candidates with additional experience in cryoultramicrotomy, confocal microscopy, and computer-aided image analysis.
Duties
All aspects of specimen preparation for a variety of biological samples. Immunofluorescence and immunogold cytochemistry. Imaging and analysis of specimens using advanced light (including confocal) and electron microscopes. Preparation of micrographs for publication. Routine maintenance of light microscopes, an ultramicrotome, and all microscopy related equipment and supplies. Aspects of experimental design and method development as commensurate with experience. Train, advise, and assist other lab members with regard to specimen preparation, microscopy, and imaging. Salary will be in the range of $ 25 - 30 K per annum, or higher, depending on experience.
Send your c.v. and the names and addresses of 3 references to:
Dr. Ann Hubbard Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine 725 N. Wolfe St. Baltimore, MD 21205
Dear Microscopists: Our imaging centre would like to acquire a 2-photon "confocal" microscope. I know of the Bio-Rad multi-photon system. Have any other companies begun to manufacture and market such systems?
David P. Bazett-Jones, Ph.D.
Professor Departments of Anatomy and Medical Biochemistry Director, Microscopy and Imaging Facility The University of Calgary 3330 Hospital Dr Calgary, AB T2N 4N1 Canada TEL: 403 220-3025, FAX: 403 270-0737
I need desplastificate slices of 4-5 um of insects legs, which are embedded in Durcupan. I have tryed with methoxide of Sodium, and saturated solution of Ethoxide of sodium ( from 1h. to 24 hrs.), and I couldn't. I have problems with the Ethoxide solution too, it become brown after two days. Somebody knows wath can I do?. Thanks, Veronica. Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
I need desplastificate slices of 4-5 um of insects legs, which are embedded in Durcupan. I have tryed with methoxide of Sodium, and saturated solution of Ethoxide of sodium ( from 1h. to 24 hrs.), and I couldn't. I have problems with the Ethoxide solution too, it become brown after two days. Somebody knows wath can I do?. Thanks, Veronica. Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
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Reply to: RE} 2-photon microscopy
Dear David, I believe Zeiss is also working on a 2-photon system.
Linda Chicoine Center for Cell Imaging http://info.med.yale.edu/cellimg Cell Biology Yale University New Haven, CT 06520 203-785-3646
--------------------------------------
Dear Microscopists: Our imaging centre would like to acquire a 2-photon "confocal" microscope. I know of the Bio-Rad multi-photon system. Have any other companies begun to manufacture and market such systems?
David P. Bazett-Jones, Ph.D.
Professor Departments of Anatomy and Medical Biochemistry Director, Microscopy and Imaging Facility The University of Calgary 3330 Hospital Dr Calgary, AB T2N 4N1 Canada TEL: 403 220-3025, FAX: 403 270-0737
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Dennis, Do you embed in Durcupan? Thanks, Veronica. Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
Hi all, A post-doc has presented me with an interesting task and I would appreciate some help! I have been given some mouse femurs (undecalcified) which were placed in a 1:1 mixture of JB-4 resin (both components A and B in the appropriate mix) and water. The samples were then placed in a vacuum at 20*C and left over the weekend with the aim of enhancing infiltration. On Monday the samples were sitting in a gelatinised resin yuk!! My task is to re-embed them. I am currently trying to dissolve the resin out with many changes of distilled water and constant movement but all that has happened is that the resin has turned white. Any advice would be welcome as these are "valuable" (aren't they all!) samples.
I need to locate any published papers on ESEM applications of petroleum technology and exploration and production in general. Any information will be greatly appreciated.
Sara, You are, as they say in the UK "in a bit of a sticky wicket". JB4 is of course GMA which in it's polymerized state is not soluable in any chemical solution that I know of. It also cannot be disolved in anything that won't also destroy your tissue samples. I know as I've worked with it for about 20 + years. Now that I've ruined you morning coffee....... try the following.
Remove ALL of the partially polymerized GMA and any water. Place in fresh GMA (no water) at 4*C for 12 hours under vacuum (preferably with agitation). Repeat this step twice more. When ready to embed the tissue allow the polymeization to occur at 4*C. Polymerization should be complete after about 4 hours. Feel free to call me at the numbers listed in my sig line at the end of this message if you have any further questions. -- Begin original message --
} Hi all, } A post-doc has presented me with an interesting task and I would } appreciate some help! I have been given some mouse femurs } (undecalcified) which were placed in a 1:1 mixture of JB-4 resin (both } components A and B in the appropriate mix) and water. The samples were } then placed in a vacuum at 20*C and left over the weekend with the aim } of enhancing infiltration. On Monday the samples were sitting in a } gelatinised resin yuk!! My task is to re-embed them. I am } currently trying to dissolve the resin out with many changes of } distilled water and constant movement but all that has happened is that } the resin has turned white. Any advice would be welcome as these are } "valuable" (aren't they all!) samples.
} Sarah Ellis } } Ps. Alcohol cannot be used. }
-- End original message --
Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chaple Hill, NC 27599 Phone 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
I am desperately looking for a used plunge freezing unit that will take liquid propane. It is for freezing algal cells. I don't want to slam them as they are spherical (even with backing I don't think this would work well....comments?).
So if ANYONE has ANY information on this could you PLEASE let me know!!!
I will be most grateful
Cheers.
Lilian Alessa Postdoctoral Fellow, Kropf Lab Department of Biology University of Utah Salt Lake City, Utah U.S.A.
I need desplastificate slices of 4-5 um of insects legs, which are embedded in Durcupan. I have tryed with methoxide of Sodium, and saturated solution of Ethoxide of sodium ( from 1h. to 24 hrs.), and I couldn't. I have problems with the Ethoxide solution too, it become brown after two days. Somebody knows wath can I do?. Thanks, Veronica. Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
Do you mean they were put into solution A plus catalyst? In any of the procedures for JB-4 that I have come across, the solution B is only added at the hardening step. The catalyst only encourages the plastic to set up in a quick and orderly manner, but the solutions A and B are the primary components of the plastic. If they are both in the mixture, eventually you can get hardening in any closed container and placing it into a vacuum only hastened the process. I'm not suprized at the reaction with water that you got. Typically, I float fresh-cut sections on water to stretch them and get them to stick to the slide. The JB-4 gets sticky, but doesn't entirely dissolve. I haven't had a block go gooy on me, but it might help if you put it in a fresh change of soltuion A without any solution B or catalyst added. I assume you have manualy removed as much JB-4 from the tissue as you can. Good Luck.
Karen Pawlowski
On Wed, 20 Aug 1997, ellis, sarah wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi all, } A post-doc has presented me with an interesting task and I would } appreciate some help! I have been given some mouse femurs } (undecalcified) which were placed in a 1:1 mixture of JB-4 resin (both } components A and B in the appropriate mix) and water. The samples were } then placed in a vacuum at 20*C and left over the weekend with the aim } of enhancing infiltration. On Monday the samples were sitting in a } gelatinised resin yuk!! My task is to re-embed them. I am } currently trying to dissolve the resin out with many changes of } distilled water and constant movement but all that has happened is that } the resin has turned white. Any advice would be welcome as these are } "valuable" (aren't they all!) samples. } } Thanks in advance, } } Sarah Ellis } } Ps. Alcohol cannot be used. }
When doing standardless quant. work on the EDX, with an accelerating voltage of 30 KeV, and getting K, L & M lines, what is the rule to follow as far as selecting lines to use in the analysis. Am I supposed to pick the ones with the most counts? Should I pick all of the same, as in Hg-K, Cu-K, Si-K etc? Should I pick the highest energy ones of those detected Because as I'm sure most of you are aware, I get completely different results when I pick the different sets of lines. Mark Darus
You can also keep your EtOH dry by using Drying Beads (Molecular Sieves). We keep ethanol in 200-500 ml bottles with the beads, and bake them out every time we use up the 200-300 ml of ethanol. This saves discarding your expensive abs. ethanol when the bottle has been open for a while. (Caution: make sure you pour out the beads into a pan and let the ethanol evaporate entirely before putting them into the oven, or they will explode and fly EVERYWHERE inside the oven.) Also keep the lid on the bottle except when actually removing some to prevent H2O condensation inside.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Digital Instruments (DI) will be hosting two free workshops on Scanning Probe Microscopy in the New England area. Those interested in attending can find further details at the DI web site www.di.com.
I realize this is only a VERY distantly related question, but I appreciate any help you can give me.
Yesterday I was asked if I know of a source of strepavidin coated microscope slides. One of the investigators I work with wants to use biotinilated cDNA probes and do some confocal microscopy on the result (apparently with other fluorescent probes). His previous attempts have had most of the DNA not stick to the slide through the entire thermocycling process so he's looking for another way to do it. Any ideas on a source for such slides?
TIA, Doug ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html
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Hi everyone,
The Webmaster of our server tells me that there have been a LARGE number of failed page requests for the Microscopy Pages that I posted last week. I'm sorry for the problems, the long URLs I have to work with apparently wrapped to a second line on many people's email readers & when they tried to access the pages the URL was missing the last few characters (computers are SO literal).
If you're still interested in checking out my series of WWW pages on topics such as general microscopy, histology, confocal, electron microscopy, digital imaging and a list of free publications that are of interest to microscopists you can get to them by this much shorter URL (the links are in the middle of this page): http://www.pharmacy.arizona.edu/exp_path.html
Thanks for your patience.
Yours, Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html
I don't know if all manufacturers have this reported in the results, but my TN Voyager gives me data for "Atom %" and "Element %", in many cases the element in majority is different. Can someone explain what these 2 mean, or at least tell me where to go to find out. Thanks
} I don't know if all manufacturers have this reported in the } results, but my TN Voyager gives me data for "Atom %" and "Element %", } in many cases the element in majority is different. Can someone explain } what these 2 mean, or at least tell me where to go to find out. ...
X-ray analysis is primarily sensitive to weight percent ... e.g., lead sulfide (PbS) Pb:S = 87:13 ... however, there is generally always a software option to cast wt% as atomic percent ... i.e., Pb:S = 50:50. Does this answer your question??
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Dear Colleagues: A technician here has difficulty to stain mouse CD4 cells by immunofluorecense using an antibody of anti-mouse CD4 conjugated with biotin, and then FITC-avidin. My suspicion is that the problem is with the antibody. As far as I know of, some antibodies are not suitable for immunocytochemistry even if then work well in other tests like ELISA or Immunoblotting. Could some of you recommend some antibody source where we can purchase such Abs against mouse CD4 and CD8 antigen that work well in immunocytochemistry? Thank you very much in advance. Regards, Yuhui Xu, MD,PhD EM Core, DFCI
Lilian, You might be interested in the paper we just published in Microscopy Research & Tech; S. D. Fields, G. W. Strout, & S. D. Russell. Spray-Freezing Freeze Substitution (SFFS) of Cell Suspensions for Improved Preservation of Ultrastructure Micros. Res. & Tech. 38:315-328 1997. The spray freezing device we've developed is decribed within. We have successfully frozen algal and other cells with this method. Greg
Lilian Alessa (by way of Nestor J. Zaluzec) wrote: } } } } Greetings All!! } } I am desperately looking for a used plunge freezing unit that will take } liquid propane. It is for freezing algal cells. I don't want to slam } them as they are spherical (even with backing I don't think this would } work well....comments?). } } So if ANYONE has ANY information on this could you PLEASE let me know!!! }
-- ======================================================== Greg Strout Electron Microscopist, University of Oklahoma e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not neccessarily those of the University of Oklahoma ========================================================
A colleague of mine (no, it was NOT me) mistakenly ran some RC prints through a heated drum dryer with predictable results: the plastic melted onto the drum. Now this person was wondering how to remove the mess. My suggestion: use water soaked towels to remove the paper part and acetone to dissolve the plastic. Any other possibilities? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
My EDX system is different, but I would assume that manufacturers pretty much use the same nomenclature and means of obtaining results. It is my understanding that "Element %" is the weight percent calculated for each element, and is not normalized to 100%. This value is a good way to check your analysis; the total % of all elements should not deviate from 100% by very much (I use + or - 2%) unless you have a problem with your analysis. "Atom %" is atomic percent, which is determined by taking the weight percent of each element and dividing by its atomic weight, normalizing, and then determining the atomic percent. This is probably why you have different results for the majority element.
Regards,
Bob ************************* Bob Citron Chiron Vision Claremont, CA USA (909) 399-1311 Bob_Citron-at-cc.chiron.com *************************
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I don't know if all manufacturers have this reported in the results, but my TN Voyager gives me data for "Atom %" and "Element %", in many cases the element in majority is different. Can someone explain what these 2 mean, or at least tell me where to go to find out. Thanks
Does anyone know the approximate amount of protein in solution which would jell a glutaraldehyde-based fixative solution? I have seen this phenomenon occasionally in the past when I was working on animal material where there was alot of cellular damage as part of the experimental design. But, now I've seen this phenomenon while preparing fruit samples (berries) and I don't quite know what to make of it, since glutaraldehyde is a protein crosslinker. We estimate the protein content of our samples to vary between 1 and 10%.
Thanks for any ideas.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Dear Mark, The general rule is to use a K line when you can, since they are the best characterized and have the best gaussian profile. L's are next and quant. analysis using M lines is usually imprecise. Of course, you must have sufficient overvoltage for the line you want to analyse, try for at least twice the line energy in your acc. voltage (hence a 10 keV x-ray range and a 20 kV acc. voltage). I have not had much luck doing standardless quant. at 30 kV and a 0 to 20 keV range. I find 20 kV and a 0 to 10 keV to be better. Do lots of work with known samples close to the content of your unknowns. BTW, you'll have trouble getting the Hg-K line, it has an energy around 78 keV. You wrote:
} When doing standardless quant. work on the EDX, with an accelerating } voltage of 30 KeV, and getting K, L & M lines, what is the rule to } follow as far as selecting lines to use in the analysis. Am I supposed } to pick the ones with the most counts? Should I pick all of the same, as } in Hg-K, Cu-K, Si-K etc? Should I pick the highest energy ones of those detected Because as I'm sure most of you are aware, I get completely different results } when I pick the different sets of lines. } Mark Darus } } G. E. Lighting Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} Date: Wed, 20 Aug 1997 22:56:24 -0700 } To: "Mark E. Darus (216) 266-2895 General Electric Co." {darus-at-cle.dnet.ge.com} } From: Mary Mager {mager-at-unixg.ubc.ca} } Subject: Re: Lines for Quant. work } Cc: Microscopy } } Dear Mark, } The general rule is to use a K line when you can, since they are the best characterized and have the best gaussian profile. L's are next and quant. analysis using M lines is usually imprecise. Of course, you must have sufficient overvoltage for the line you want to analyse, try for at least twice the line energy in your acc. voltage (hence a 10 keV x-ray range and a 20 kV acc. voltage). I have not had much luck doing standardless quant. at 30 kV and a 0 to 20 keV range. I find 20 kV and a 0 to 10 keV to be better. Do lots of work with known samples close to the content of your unknowns. BTW, you'll have trouble getting the Hg-K line, it has an energy around 78 keV. } You wrote: } } } When doing standardless quant. work on the EDX, with an accelerating } } voltage of 30 KeV, and getting K, L & M lines, what is the rule to } } follow as far as selecting lines to use in the analysis. Am I supposed } } to pick the ones with the most counts? Should I pick all of the same, as } } in Hg-K, Cu-K, Si-K etc? Should I pick the highest energy ones of those detected Because as I'm sure most of you are aware, I get completely different results } } when I pick the different sets of lines. } } Mark Darus } } } } G. E. Lighting } Regards, } Mary } } Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Hi We do a lot of diffraction pattern interpretation. Measuring the patters are a tedious job, and in polycrystalline materials it is difficult to measure ring patterns accurately. I have a few questions: *Is there any software available to help this process? That is measuring 'a' as well as the option to try fit the values to standard elemental values from the JCPDS tables. *Does new TEM's (eg Philips CM series) have a measuring option in their software? * If such programs exists, what's the accuracy/resolution of it?
Thanx Sara
-------------------------------------------------------------------------------------- Sara Prins Surface and Structure Analytical Services Division for Materials Science and Technology CSIR PO Box 395 Pretoria South Africa Tel: +27+12+8413974 Fax: +27+12+8414395 sprins-at-csir.co.za Visit us at : http://www.csir.co.za
One problem associated with Molecular Sieve in ethanol is the possibility of dust from the sieve being released into the ethanol. This dust then sticks to the specimen and eventually damages the knife. We have stopped using a drying agent in the ethanol, rather decanting ethanol into smaller (250ml) containers and replacing at regular intervals - saves on diamond knife resharpening!
} } You can also keep your EtOH dry by using Drying Beads (Molecular } Sieves). We keep ethanol in 200-500 ml bottles with the beads, and bake } them out every time we use up the 200-300 ml of ethanol. This saves } discarding your expensive abs. ethanol when the bottle has been open for } a while. (Caution: make sure you pour out the beads into a pan and let } the ethanol evaporate entirely before putting them into the oven, or they } will explode and fly EVERYWHERE inside the oven.) Also keep the lid on } the bottle except when actually removing some to prevent H2O condensation } inside. } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735 }
Prof Jan Coetzee Head: Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/science/electron/emunit1.htm
} One problem associated with Molecular Sieve in ethanol is the } possibility of dust from the sieve being released into the ethanol. } This dust then sticks to the specimen and eventually damages the } knife. We have stopped using a drying agent in the ethanol, rather } decanting ethanol into smaller (250ml) containers and replacing at } regular intervals - saves on diamond knife resharpening!
In order to eliminate the dust problem we place the molecular sieve in a small piece of dialysis tube that is closed using metal clips. We have no water and no dust in acetone, methanol, ethanol. An other way to get rid of water in solvents is to add a small amount of acidified 2,2-Dimethoxypropane. The DMP will react with water to produce ethanol and acetone. If you do not mind small amounts of acetone and DMP in your ethanol this is a very simple way to dry it.
A first guess might be that you've got pectin - of jam and wine haze fame. If the berries make good jam that might be the answer. Malcolm Haswell e.m unit University of Sunderland UK ----------
Hello, everyone,
Does anyone know the approximate amount of protein in solution which would jell a glutaraldehyde-based fixative solution? I have seen this phenomenon occasionally in the past when I was working on animal material where there was alot of cellular damage as part of the experimental design. But, now I've seen this phenomenon while preparing fruit samples (berries) and I don't quite know what to make of it, since glutaraldehyde is a protein crosslinker. We estimate the protein content of our samples to vary between 1 and 10%.
Thanks for any ideas.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
} One problem associated with Molecular Sieve in ethanol is the } possibility of dust from the sieve being released into the ethanol. } This dust then sticks to the specimen and eventually damages the } knife. We have stopped using a drying agent in the ethanol, rather } decanting ethanol into smaller (250ml) containers and replacing at } regular intervals - saves on diamond knife resharpening!
This discussion has been brought up previously on the list. Many labs encase the molecular sieves in dialysis tubing to prevent the dust from contaminating the solution.
Dennis Shubitowski University of Michigan dshubito-at-umich.edu
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Memo : job opp.: intermetallics 21-08-1997 15:48
Hi all,
We're looking for a TEM specialist with practical knowledge of intermetallics for a 6 or 12 month (Jan. - Dec. 1998) post-doc position at the Electron Microscopy for Materials Science group in Antwerp, Belgium, and working on Ni- and Fe-based materials.
If you're interested please contact me as soon as possible. The application can only be started when the applicant is known.
Thanks,
Nick Schryvers
Dr. Dominique Schryvers University of Antwerp, RUCA - EMAT Groenenborgerlaan 171, B-2020 Antwerpen (Belgium) Tel: 32-3-2180247 Fax: 32-3-2180257 e-mail: schryver-at-ruca.ua.ac.be homepage: http://www.ruca.ua.ac.be/~EMAT/Schryvers.html
The rule of thumb to follow if possible use the lines in order K, then L then M. If using standardless analysis you want to use the lines in the same family if possible preferably all K lines. Be careful of overvoltage if using a 30kV beam for some of the lower energy lines. Is 30kV necessary to excite all of the lines that you want? The rule of thumb is to use at least twice the energy of the line as your accelerating voltage.
My experience with standardless analysis has also been that there is less acuracy with increaseing elements. Brasses and Bronzes come up very well but the accuracy trails off as you add more elements. And super alloys are almost impossible. I hope that this has been hepful.
As to the question of weight percent and atom percent. The atom percent is basicaly what percentage of atoms of one element there are in relation to another (ie a 1:1 ratio of Cu to Zn would produce a 50% at% Cu and 50 at% Zn where a 7:3 ratio would produce a 70 at% Cu to 30 at% Zn). The weight percent takes into account the atomic number of the elements or their atomic weight therfore a 50 at% Al and 50 at% W would show a weight percent much higher for the W atom since it is much heavier. I hope this has been helpful. Please let me know if you have anymore questions.
__________________________ Roberto Garcia EMF Manager Wright State University rgarcia-at-cs.wright.edu
I have to say that when I have seen light/dark bands on slow scan (mainly textiles) I usually find that the problem is charging on the sample--not a microscope problem. I believe it to be a capacitance effect where the sample charges and dissipates. Usually, recoating the sample with gold or using Fullam's antistatic liquid (I hang my head in shame!) solves the problem.
Changing the voltage may help troubleshoot the problem as well.
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Olli,
They kept trying to say that my problems were fields, but the active field cancellation could not affect the visual symptoms. The ultimate proof that it was inside the system, is the fact that the problems went away when they replaced the entire column. It is easy for SEM manufacturers to blame fields for problems they can not trace, or easily solve. They all seem to use this crutch from time to time.
Do not get me wrong, I know fields can be a problem. My response to that is that, as the systems become more and more susceptible to the ocean of magnetic noise they live in, the manufacturers need to prevent the affects with their design, and not use it as an excuse for poor performance in the real world.
Enough of that! I was just wondering if anyone else was having these problems on similar systems. It seems as though there are a few out there.
I wanted to find out if anyone could tell me if they use LR White as an embedding resin NOT for immuno work. The reason why I say not for immuno work, is that I would like to osmicate them. I was curious to see if I fix samples in Gluteraldehyde, and then osmicate them, could I embed them up in LR white with Gelatin capsules/coverslips and then section and stain for the TEM. How does this resin hold up under the beam? I haven't seen any one mention this on this listserver, and wondered if people even do this. Generally, I use Spurrs resin, but have some extra LR White resin that I would like to use up before it expires.
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
Just wanted to report that using a fresh bottle of absolute ETOH has solved the "pinhole" artifact ... such an easy fix for such an irritating problem. Thanks again for your help. (The additional conversation about the sieves has also been enlightening.) Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
In message {s3fc16e2.081-at-EM.AGR.CA} Susan Carbyn writes:
} I wanted to find out if anyone could tell me if they use LR White as an } embedding resin NOT for immuno work. The reason why I say not for } immuno work, is that I would like to osmicate them. I was curious to see } if I fix samples in Gluteraldehyde, and then osmicate them, could I embed } them up in LR white with Gelatin capsules/coverslips and then section } and stain for the TEM.[?]
Sure, and because of its very low viscosity, LR White works well for infiltrating into plant tissues.
} How does this resin hold up under the beam?
Not as well as epoxy sections do, on uncoated grids. They can drift, tear or flap in the electron "breeze". Solution: 1. use Formvar or similarly coated grids to stabilize the sections. 2. Coat LR White sections (mounted on bare grids) with thin carbon layer in a vacuum evaporator, taking care to minimize heat delivered to sections. 3. Use higher mesh grids, eg. 200-400#.
} Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } E-mail: carbyns-at-em.agr.ca } } Phone: (902) 679-5566 } Fax: (902) 679-2311
Good luck!
Gib
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
LR White is a good embedding medium for staright morphology after osmication. Doesn't always section as nicely as an epoxy, but still very useful for hard to embed materials. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 10:18 AM 8/21/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Susan Carbyn wrote: } } Just a quick question: } } I wanted to find out if anyone could tell me if they use LR White as an } embedding resin NOT for immuno work. The reason why I say not for } immuno work, is that I would like to osmicate them. I was curious to see } if I fix samples in Gluteraldehyde, and then osmicate them, could I embed } them up in LR white with Gelatin capsules/coverslips and then section } and stain for the TEM. How does this resin hold up under the beam? I } haven't seen any one mention this on this listserver, and wondered if } people even do this. Generally, I use Spurrs resin, but have some extra } LR White resin that I would like to use up before it expires. } } Susan, You can use the LR White as long as you cure the resin in the oven. The Osmium will not cause any problems. The resin holds up fine in the beam. Just remember to exclude exposure to oxygen or the resin will not cure.
Greg Rudomen Greg-at-umic.sunysb.edu University Microscopy Imaging Center SUNY Stony Brook
I had some misfortunes with osmium-fixed tissue embedded in LR White resin. The resin polymerized prematurely during infiltration, but it worked well with glut-fixed tissues. Someone thought that I had too much an accelerator. The real problem was that LR White was too old and it reacted with osmium. A new bottle of resin worked well for a while and then it acted up again.
I think the way to counter this problem is to buy LR White without the accelerator already mixed. When needed, one can mix them up and divide into several portions for storage. A portion will be warmed up each time and it can be used up quickly. The rest of the resin stays cold, therefore, it can be kept for a long time without causing problems.
Ann Fook Yang, EM Unit, ECORC, Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada K1A 0C6
} } } Susan Carbyn {CarbynS-at-em.agr.ca} 08/21/97 10:18am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just a quick question:
I wanted to find out if anyone could tell me if they use LR White as an embedding resin NOT for immuno work. The reason why I say not for immuno work, is that I would like to osmicate them. I was curious to see if I fix samples in Gluteraldehyde, and then osmicate them, could I embed them up in LR white with Gelatin capsules/coverslips and then section and stain for the TEM. How does this resin hold up under the beam? I haven't seen any one mention this on this listserver, and wondered if people even do this. Generally, I use Spurrs resin, but have some extra LR White resin that I would like to use up before it expires.
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
We use LR White for non-immuno routinely. We use grids without = supporting membranes for both LR White and Embed. It holds up in the = beam, although is a bit less stable than Embed or any of the other Epon = replacements and is only a problem with some of the newest students who = let a crossover beam sit on it. Gelatin capsules can be used and some people also use BEEM capsules. = Some of the BEEM type capsules however are more permiable to oxygen, but = others seem to work OK. It is important that all the EtOH be removed = before embedding. As such we do not use EtOH:resin, 2:1, 1:1, and 1:2 as = we do in Embed. We go through 100% EtOH twice, then into 3 changes of = pure LR White. We love it and all the students love it. I personally have used LR White = since before it was actually an EM product because it is so fast and easy = to use. Hope comments are helpful. Judy M.
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
Phone: 209/954-5284 FAX: 209/954-5600 e-mail: jmurphy-at-sjdccd.cc.ca.us program web page: http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html
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Just a quick question:
I wanted to find out if anyone could tell me if they use LR White as an embedding resin NOT for immuno work. The reason why I say not for immuno work, is that I would like to osmicate them. I was curious to see if I fix samples in Gluteraldehyde, and then osmicate them, could I embed them up in LR white with Gelatin capsules/coverslips and then section and stain for the TEM. How does this resin hold up under the beam? I haven't seen any one mention this on this listserver, and wondered if people even do this. Generally, I use Spurrs resin, but have some extra LR White resin that I would like to use up before it expires.
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
E-mail: carbyns-at-em.agr.ca
Phone: (902) 679-5566 Fax: (902) 679-2311
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;21 Aug 1997 10:28:24 -0800 Received: from Sparc5.Microscopy.Com (206.69.208.10) by = ms.sjdccd.cc.ca.us with SMTP (Apple Internet Mail Server 1.1.1); Thu, 21 Aug 1997 10:28:12 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id JAA02109 for dist-Microscopy; Thu, 21 Aug 1997 = 09:20:01 -0500 Received: from agrout1.agr.ca (agrout1.agr.ca [192.197.71.131]) by = Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id JAA02106 for = {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Aug 1997 09:19:59 -0500 Received: from agrgate.agr.ca (agrgate.agr.ca [192.197.71.189]) by agrout1.agr.ca (8.8.4/8.8.4) with SMTP id KAA23603 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Aug 1997 = 10:22:58 -0400 (EDT) Received: from agrin1.agr.ca by agrgate.agr.ca via smtpd (for agrout1.agr.ca [192.197.71.131]) with SMTP; 21 = Aug 1997 14:31:23 UT Received: from EM.AGR.CA (gwiseweb.agr.ca [142.61.33.38]) by agrin1.agr.ca (8.8.4/8.8.4) with SMTP id KAA24906 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Aug 1997 = 10:24:37 -0400 (EDT) Received: from AGCAN-Message_Server by EM.AGR.CA with Novell_GroupWise; Thu, 21 Aug 1997 10:22:26 -0400 Message-Id: {s3fc16e2.081-at-EM.AGR.CA} X-Mailer: Novell GroupWise 4.1
Hi,
LR White is an acrylic embedding medium. It has many advantages for immuno work, among them its low crosslinkage. It does not bind with the tissue (like epoxies) but through tissue. It does not preserve tissue as well as epoxy. It is not as beam stable as epoxy. Its polymerization reaction is exothermic - if uncontrolled - it may damage tissue. Thick sections may wrinkle badly (due to the lack of crosslinkage). Simply to use LR White because it is in the refrigerator is not a good idea. For non-immuno work it is far more advatageous to use epoxy monomers. Bye, Hildy
A quick note of congratulations on the success of the 2-Photon seminar at last week's Microscopy & Microanalysis meeting. I only had a chance to visit for the last afternoon, but it was very clear that the sessions were well attended and that the vendors got a lot of opportunity to show off their new gear and run samples.
Is there a way that those of us who could not attend the regular workshop could get a copy of the notes? Please post info on both listservers so that a wider audience can respond.
Thanks! Barbara Foster Microscopy/Microscopy Education
I am in search of the Representative on Reichert-Jung Miccrotomes in the Seattle area. We are interested in looking at a reichert ultracut microtome?? anyone with the address or phone number wold be appreciated
Eric A.Rosen Fred Hutchinson Cancer Research Center
This is a request for information from you LM types out there. Does anyone have any suggestions for the conversion from polaroid to digital outlined below? I'm sure this has been discussed but I probably ignored it since I don't deal with LM much, but I told these people I'd send this message out for them. The $15k has to cover the computer as well. Bruce said he has information on a Polaroid system and a Leco setup. Any others? We don't have a great preference between Mac and Wintel systems as we run both.
I'd appreciate any responses. Thanks in advance for the help.
Cheers,
John Vetrano Materials Interfaces Group Pacific Northwest National Laboratory _______________________________________________________________________________
John, we have $15k to purchase CCD camera for the Zeiss and Olympus microscope in metallography. We are looking to replace the polaroid camera system in place with a CCD camera. Both scopes have C-mounting capability. Will need a CCD camera for high quality metallograpy work. We would like to capture the image and store the image for future retrieval, we would like a similar system like the SEM (Gatan DigiScan). Thanks for offering to place a ad on the server list.
On Thu, 21 Aug 1997, Jan Coetzee EM Univ Pretoria wrote:
} Date: Thu, 21 Aug 1997 10:50:16 CAT-2 } From: Jan Coetzee EM Univ Pretoria {janc-at-ccnet.up.ac.za} } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: TEM Artifact } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } One problem associated with Molecular Sieve in ethanol is the } possibility of dust from the sieve being released into the ethanol. } This dust then sticks to the specimen and eventually damages the } knife. We have stopped using a drying agent in the ethanol, rather } decanting ethanol into smaller (250ml) containers and replacing at } regular intervals - saves on diamond knife resharpening! } JUST LET THE BOTTLE SIT, AND THE DUST WILL SETTLE TO THE BOTTOM. DON'T DISTURB IT WHEN REMOVING SOME AND TAKE IT OFF THE UPPER LAYER. WE NEVER HAVE ANY PROBLEMS WITH DIRTY SAMPLES OR SHORTENED DIAMOND KNIFE LIFE.
AS WITH ANY REAGENT, IF IT LOOKS CLOUDY, DISCOLORED, OR UNUSUAL, I DON'T USE IT.
S. MILLER
} } } } You can also keep your EtOH dry by using Drying Beads (Molecular } } Sieves). We keep ethanol in 200-500 ml bottles with the beads, and bake } } them out every time we use up the 200-300 ml of ethanol. This saves } } discarding your expensive abs. ethanol when the bottle has been open for } } a while. (Caution: make sure you pour out the beads into a pan and let } } the ethanol evaporate entirely before putting them into the oven, or they } } will explode and fly EVERYWHERE inside the oven.) Also keep the lid on } } the bottle except when actually removing some to prevent H2O condensation } } inside. } } } } Sara E. Miller, Ph. D. } } P. O. Box 3020 } } Duke University Medical Center } } Durham, NC 27710 } } Ph: 919 684-3452 } } FAX: 919 684-8735 } } } } } } Prof Jan Coetzee } Head: Unit for Electron Microscopy Tel:+27-12-420-2075 } University of Pretoria Fax:+27-12-342-1738 } Pretoria 0002 Internet:janc-at-ccnet.up.ac.za } South Africa http://www.up.ac.za/science/electron/emunit1.htm }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} --Paula, Not 5 minutes before your e-mail I was asked to look at some artery from a goat which was showing exactly what you described. The vessel had been incubated with Clostridium perfrinogen toxin,it was covered with a gelatenous substance. But I cant help you answer your question.
Christine Lee, Vet. Pathobiology, University of Queensland. -
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Eric, Reichert -Jung is now called Leica their # is 800-248-0123. -- Begin original message --
} From: Eric Rosen {erosen-at-fred.fhcrc.org} } Date: Thu, 21 Aug 1997 13:33:45 -0700 (PDT) } Subject: Microtome manufacturer address } To: Microscopy-at-Sparc5.Microscopy.Com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } I am in search of the Representative on Reichert-Jung Miccrotomes in the } Seattle area. We are interested in looking at a reichert ultracut } microtome?? anyone with the address or phone number wold be appreciated } } Eric A.Rosen } Fred Hutchinson Cancer Research Center } } } }
-- End original message --
Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chaple Hill, NC 27599 Phone 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
Ann-Fook Yang (Ann-Fook Yang) wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I had some misfortunes with osmium-fixed tissue } embedded in LR White resin. The resin } polymerized prematurely during infiltration, but it } worked well with glut-fixed tissues. Someone } thought that I had too much an accelerator. The } real problem was that LR White was too old and it } reacted with osmium. A new bottle of resin worked } well for a while and then it acted up again. } } I think the way to counter this problem is to buy LR } White without the accelerator already mixed. } When needed, one can mix them up and divide into } several portions for storage. A portion will be } warmed up each time and it can be used up } quickly. The rest of the resin stays cold, therefore, } it can be kept for a long time without causing } problems. } } Ann Fook Yang, } EM Unit, } ECORC, Agriculture and Agri-Food Canada, } Ottawa, Ontario, Canada } K1A 0C6 } } } } } Susan Carbyn {CarbynS-at-em.agr.ca} } 08/21/97 10:18am } } } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Just a quick question: } } I wanted to find out if anyone could tell me if they } use LR White as an } embedding resin NOT for immuno work. The } reason why I say not for } immuno work, is that I would like to osmicate them. } I was curious to see } if I fix samples in Gluteraldehyde, and then } osmicate them, could I embed } them up in LR white with Gelatin } capsules/coverslips and then section } and stain for the TEM. How does this resin hold up } under the beam? I } haven't seen any one mention this on this listserver, } and wondered if } people even do this. Generally, I use Spurrs resin, } but have some extra } LR White resin that I would like to use up before it } expires. } } Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } E-mail: carbyns-at-em.agr.ca } } Phone: (902) 679-5566 } Fax: (902) 679-2311
Dear Ann-Fook Yang, We agree with you. Ladd sells LR White and about two years ago we stopped selling it with the accelerator already mixed in for the reason you stated. John Arnott
In response to a question on glutaraldehyde, the name of the Ladd Research's head chemist, Dr. Charles Duvic was indvertently entered as "Garber". Ladd apologizes profusely to those who contacted us concerning this error. Dr. Duvic has worked for many years to develop the quality and reputation of Ladd's glutaraldehyde and other chemicals, so it is no way an insult to have ones name substituted for his. Never the less we do apologize to any one who was offended.
Information regarding the International EM Congress next year may be found at:
http://icem.inin.mx
Take a look ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
I, too, had an incident where my osmium-fixed tissue caused the LR White to polymerize during an infiltration step. I was also infiltrating samples of the same tissue without osmium and had no early polymerization problems. We have not had any other problems with LR White aside from the occasional oxygen inhibiting polymerization a bit, but we don't usually osmicate samples for LR White embedding. I did not add accelerator at any point during the infiltration and all of the samples were at the same temperature. I have not had the time to follow-up on the issue, yet.
Gregg Sobocinski Parke-Davis Pharmaceutical Research Division Ann Arbor, Michigan, USA Sobocig-at-aa.wl.com -------------------------------------------------------------------. } } I had some misfortunes with osmium-fixed tissue } embedded in LR White resin. The resin } polymerized prematurely during infiltration, but it } worked well with glut-fixed tissues. Someone } thought that I had too much an accelerator. The } real problem was that LR White was too old and it } reacted with osmium. A new bottle of resin worked } well for a while and then it acted up again. } } I think the way to counter this problem is to buy LR } White without the accelerator already mixed. } When needed, one can mix them up and divide into } several portions for storage. A portion will be } warmed up each time and it can be used up } quickly. The rest of the resin stays cold, therefore, } it can be kept for a long time without causing } problems. } } Ann Fook Yang, } EM Unit, } ECORC, Agriculture and Agri-Food Canada, } Ottawa, Ontario, Canada } K1A 0C6
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} ...Gelatin capsules can be used and some people also use BEEM capsules. } Some } of the BEEM type capsules however are more permiable to oxygen, but } others seem } to work OK...
Flat molds for specimen orientation can be a problem, since most are permeable to LR White. Pella sells a teflon flat mold that solves the problem.
} From Susan Carbyn:
} I wanted to find out if anyone could tell me if they use LR White as an } embedding resin NOT for immuno work.
LR White Hard grade is great for hard biological (bone, keratinized epithelium, etc.) and non-biological (catalysts, hard polymers, etc.) samples.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Hi all, I'm looking for the power supply for a Zeiss Photomicroscope III (Zeiss p/n 47 20 83). If you have one for sale, trade, or donation, please contact me at the address below. TIA Julian
Julian P.S. Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x227 (vox) 803-323-2246 (fax)
Does anyone know of any sources for antibodies against phosphorylated forms of MAP-2 (microtubule associated protein-2)?
Thanks in advance, Glen MacDonald Virginia Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu *---------------------------------------------------------------------* The box said "Requires Windows 95 or better.", so I bought a Macintosh. *---------------------------------------------------------------------*
Thanks to the ten or so people who replied to my question about bakeout frequency and flash intensity on Hitachi S4500 FESEMs, either directly or via the server. The word ' from the horse's mouth' was that the flash current should be around 25-30uA for a clean tip - higher may indeed damage the tip, and bakeout should be done when the vacuum deteriorates, inter-flash interval decreases too much, or tip noise increases and cant be decreased by running at 30kV for 2-4 hours then 5kV. The range of replies was awesome - one person has been flashing in the forties for years, bakeout recommendations varied from monthly to "not done in 60 months" - but everyone seemed happy with the performance under the regime they were using.
So, we baked out, adjusted the flash intensity (call your service man), and made good resolutions about keeping records of flash current and post flash extraction voltage. And always using the cold trap.
For the record, we are a service unit with a wide range of users, we have a diffusion-pumped chamber and run a cold stage from time to time. } } Hi all, } I am after some advice on the care of the FE tip and vacuum } on a Hitachi 4500. } } 1. How often should one bake out? The manual recommends baking out } when the vacuum deteriorates. After about 8 months operation ours is } better than it was to start with - IP1&2 off-scale, IP3 at } 7x10-7 Pa. On the other hand many people seem to recommend baking at } fairly short intervals "whether it needs it or not". We are inclined } to a non-interventionist approach but are getting a bit nervous...any } advice? } } 2. What should the flash current intensity be? Ours started at around } 15-20 (and we sometimes flashed twice to get a reading in the high } twenties) but has steadily crept up and is now in the high forties. } Is this good, bad or indifferent? Is it perhaps related to question } 1? If it gets too high does it wreck the tip? We are generally } flashing once or twice a day. ---------------------------------------------------------------------- Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 6 249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
If you are (*were!*) a member of Histonet, please note the server was disabled by a lightning strike and power outage. The power problems also took out the backup system!
Herb Hagler informs us that things are now back to normal, but former members will have to resubscribe to the listserver.
Send a message to: Histonet-at-pathology.swmed.edu
with the following as the subject: subscribe
Please pass this information to your friends and colleagues who were subscribers to Histonet.
Thank you for informing me about your product but i have a free copy of Evex's Analytical copy of VIDX Microanalysis software. The fellows there have already confiqured my first system for me and they are working to get the second system up and running.
They were able to use my old Kevex pulse processor and power supply and interface directly to a PC using windows 95 or NT. It want even that expensive. I was able to get a demoe unit for less than $10,000.00.
Cheers Craig Ross
purchased the VIDX softwaare Buying software to work with my old equipment does not chnage the fact that the old equipment can still break and that in the long run I might be better off just buying an Evex System. It
. Fortunatelky there are independent organization such as Evex Analytical that service old equipment like my Kevex and other equipment such as Tracors and PGT's that help keep it alive , But Is the software your selling
Personally, I dont believe anything from a person or company that does not sign their name.
My Two Cents Bill Zender
--------------------- Forwarded message:
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Craig Ross wrote: "I am search for old Kevex parts for Kevex 7000 system. Our has bit the dust and we dont have too much money to spend on a new system." Dear Craig, It is very difficult to maintain old EDX systems and new ones are often prohibitively expensive. If your pulse processor is still working, consider upgrading your system to Microsoft Windows as offered by our company and several others. For a free demo software check out our website. Mektech Inc. www.visionol.net/~mektech
Bill Zender wrote: } Personally, I dont believe anything from a person or company that does not } sign their name.
} My Two Cents } Bill Zender
Dear Bill,
It's our company policy to sign with the company's name when mentioning our products on the listserver. This leaves no doubt as to our interests and is far more honest than posing as "EDXUSER"
Today there are many alternatives for the user who has a good detector but an ageing set of electronics.
Just keep the detector: If the detector is still good, one can keep it and replace the bias supply, pulse processor and MCA. A complete replacement (not including a PC of the user's choice) system including quantitative analysis software is available from ANS for $13,990. Replacing the entire electronics package can often improve both the resolution and count rate performance of a system.
Keep the detector, bias supply and pulse processor: Upgrade consists of a new MCA and Windows software. In this case ANS's upgrades can run from $4400 for a semi-Q package to $8,990 for a fully quantitative system (PC supplied by user).
Various companies have different approaches to the upgrade issue. We would recommend that anyone who is considering an upgrade should check out all of the possibilities and download evaluation software (ANS's is available at www.ansxray.com) whenever possible.
Bill Hardy President American Nuclear Systems, Inc. Manufacturer of EDS upgrade packages
Dear Listserver Readers, This is a commercial posting by Thomson Scientific Instruments Pty. Ltd. Whilst I absolutely abhor the practice of commercial postings, the recent thread concerning EDX upgrades is an obvious setup and commercially I am left with no option but to respond.
Given that our competitors have been advertising their wares I would like to point out that we also produce a upgrade package for old EDS systems and have done since 1993.
With sincere apologies to any readers who consider this posting inappropriate,
Paul Thomson Technical Director Thomson Scientific Instruments Australia Web Page: http://werple.net.au/~tsi/
This is written in response to Paul Thomson's posting. While his claim that the recent discussions have been a jackup may be true (if so it went over my head), I have welcomed the contributions from the vendors, including Paul's, as knowledge of what's currently available is always welcome in my book, as long as it's not rammed down my throat. In fact, I would like to invite vendors of upgrade equipment to contact me, as my beloved Link QX2000 is, I'm afraid, unlikely to make it into the millenium, and I'm unlikely to be able to afford a new Oxford system. I want full quantitative analysis and quantitative mapping (preferably real-time), as it will be for an EDS-only EPMA (now there's what some may regard as an oxymoron). Maybe you'd better contact me directly.
thanks
Ritchie
ps does anyone know if anybody has set up a listserver for XRF yet?
cheers
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
There has been a rash of postings the last few days about X-ray Analysis Systems. I appreciate the fact that some of our subscribers are manufacturers and whole heartily support their participation on the Listserver...
However, as you will all recall, one of our cardinal rules is no advertising. This last round of postings started by one vendor and then followed on by a number of others has begun to cross that line. Please, keep you postings to answering questions.
For example, recommendations on keeping a detector and just replacing the electronics are fine. Or stating that you manufacture a product which solves this problem also okay but then direct the reader to your WWW site for details.
However, I do not wish to see items posting that say
......... and for $xxx.yy we can sell you a product that does .........
that type of posting is crossing the line, which I admit is gray, but nevertheless is against the philosophy of this listserver. That is clearly selling rather than giving information.
There have been a number of manufacturers that have done this recently, and a number that have complained to me privately. As many of you know I usually only send out private messages to the company that crosses the line, however in this case I see the potential for many to say , well , I'll do it too. Please refrain from this type of posting.
Thanks
Nestor Your Friendly Neighborhood SysOp/Policeman?
Exerpt from the RULES of the LISTSERVER......
---------------------------------------------------------------------------- Can I post an Advertisement? ----------------------------------------------------------------------------
No, that does not fit within the bounds of this discussion forum.
This listserver is not intended to be a Sales mechanism for commerical organizations, but rather it is an open discussion area about microscopy and microanalysis problems and solutions. If you are an organization and have equipment you wish to donate, or sell, for nominal cost (i.e. no profit) then this is generally an acceptable posting. If you are not sure then send a copy of the announcement in question to Zaluzec-at-MSA.Microscopy.Com and I will give you my opinion. An example of this type would be an old decommissioned instrument which someone is trying to give away for removal/shipping costs, that would fit within the bounds of the purposes of this list.
If you are a manufacturer, you are always welcome to observe/join in any discussion at any times. We do ask that everyone, please refrain from overt sales pitches and/or commericalism. If a product which you produce/sell can solve a problem or answer a question raised by anyone on this list, then by all means feel free to say so. Try to be brief about the product, state the simple facts in a few (short) sentences and then offer to continue the discussion with any interested parties offline. Alternatively you can give sufficient information so that individuals can download/access the relevant information.Usually it will be sufficient to just add your phone number and/or Email address to the end of your message, and you'll be contacted by anyone that is interested.
Remember, please keep your comments about any product you "sell" to a minimum.
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If you are interested in using the Internet for Commerical Advertising of Microscopy/Microanalysis Related Products/Services, you may wish to contact MSA at it's WWW site (http://www.msa.microscopy.com) or the MSA Business Office (MSABusinessOffice-at-MSA.Microscopy.Com). These alternative Internet services, are provided independently of the Listserver Operation, which MSA provides as a FREE service to the WorldWide Microscopy and Microanalysis Community. Any funds derived from the above are used to defray the costs of running MSA's Internet site.
--IMA.Boundary.511815278 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
John,
Are you referring to a ferrotyping drum drier? If so, GOOD LUCK at not scratching the surface. I would think that a soft cotton towel (not paper) soaked in a solution of water and wetting agent to remove the paper residue but also make sure that the paper doesn't rub the surface as it comes off. I really think that you better call the paper manufacturer and get their suggestions on removing the coating material.
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A colleague of mine (no, it was NOT me) mistakenly ran some RC prints through a heated drum dryer with predictable results: the plastic melted onto the drum. Now this person was wondering how to remove the mess. My suggestion: use water soaked towels to remove the paper part and acetone to dissolve the plastic. Any other possibilities? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
(IMA Internet Exchange 2.1 Enterprise) id 0018E94F; Fri, 22 Aug 97 21:01:57 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ns2.baxter.com (8.8.0/8.8.0) with SMTP id VAA04630; Fri, 22 Aug 1997 21:09:33 -0500 (CDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id QAA16145 for dist-Microscopy; Wed, 20 Aug 1997 16:38:58 -0500 Received: from saluki-mail.siu.edu (saluki-mail.siu.edu [131.230.252.17]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id QAA16142 for {microscopy-at-sparc5.microscopy.com} ; Wed, 20 Aug 1997 16:38:57 -0500 Received: from [131.230.97.71] (mac71.chem.siu.edu [131.230.97.71]) by saluki-mail.siu.edu (AIX4.2/UCB 8.7/8.7) with SMTP id QAA10624 for {microscopy-at-msa.microscopy.com} ; Wed, 20 Aug 1997 16:45:52 -0500 (CDT) X-Sender: bozzola-at-saluki-mail.fiber2.siu.edu Message-Id: {v02130502b021154f05c1-at-[131.230.97.71]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I would appreciate comments from anyone using PhotoShop on a Unix based system. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Hi, Can anyone help me for the following problem : I need to visualize waxes on plant cuticles in TEM Are there methods to contrast waxes by chemicals like OSO4 for lipids for instance. I looked for some informations in literature but didn't find something interesting... Lack of time... I need to know about it so quickly as possible Any informations are welcome Thanks to All Pascal
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
I believe that there has been mention of an atomic force microscopy listserver at some time on this list. I am at home, recovering from surgery and can't look through my filed mail at the office.
We have begun doing some AFM work and I would like the address of such a server, if it does exist. I'd like to make some good use of my convalescence and learn more about the AFM before I return for work.
Bob Lawrence
"The valley of the spirit never dies; It is the woman, primal mother. Her gateway is the root of heaven and earth. It is like a veil barely seen. Use it; it will never fail."
---------------------------- Forwarded with Changes ---------------------------
Position : Analytical Equipment Engineer ---------------------------------------- Requirement : ------------ - Proficient in both practical and theoretical aspects of Secondary, Backscatter and EDS for Field Emission Scanning Electron Microscopy. - Experience in various Electron Optic equipments (FESEM, Auger and TEM) - Knowledgable in ion beam type equipment (Focus Ion Beam) - Familiar with semiconductor fabrication process and material science. - Bachelor/Master/Phd Degree in Material Science or Electrical Engineering - Good technical skill in beam based FA equipment (SEM / EDX, FIB, or TEM), material science or semiconductor fabrication process
Job Function ------------ - Be a part of Intel's dynamic and technical team performing failure analysis on Intel Microprocessors - Improve product yield, quality and reliability through in-depth failure analysis to identify defect using state of the art FA equipments (SEM/EDX, FIB) and through detailed understanding of fabrication technology - Opportunity to work very closely on Intel advanced multilayered fabrication processes (0.40 um and 0.25 um process technology) - Involved in supporting new product transfer and startup, automate and improve the failure analysis process and proliferate shared learning across Intel sites. - Job requires the condidate to be PERMANTLY STATIONED in INTEL PENANG, MALAYSIA
Candidates please contact:
In Malaysia:
Eng Hong Yeoh Tel: 604-859-6160 FAX: 604-859-6749 e-mail: Eng_Hong_Yeoh-at-ccm.ipn.intel.com
or in the US:
Kian Sin Sim, John Mardinly or David Susnitzky Intel SC2-24 2200 Mission College Blvd. Santa Clara, CA 95052-8119
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com =
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
Welcome to the Scanning Probe Microscope List/Digest!
This list is intended to be a forum for discussing views, issues, and applications of Scanning Probe Microscopy with the goal of expanding knowledge of SPM and bringing the SPM community closer together. =
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Just to add my own $0.02 worth here, I have also experienced this problem. At that time I was infiltrating plant tissue, so it needed a long time to infiltrate because of cell walls, but the LR White seemed very non-viscous (what IS the word for non-viscous, I'm always looking for that word), very much like water, and then, with no accelerator it suddenly polymerized in the vials and surprized me and caused no end of grief because of that surprize. I thought that it would slowly increase in viscosity, such that I would be able to predict when it was going to polymerize, and this is what fooled me. It behaved quite differently from other resins that I've used in the past.
This was at least 6 years ago, so I cannot remember now if I had osmicated that tissue or not.
Garry
} ---------- } From: SOBOCIG[SMTP:sobocig-at-aa.wl.com] } Sent: 22 August, 1997 07:04 } To: Ann-Fook Yang; CarbynS-at-em.agr.ca; Microscopy-at-Sparc5.Microscopy.Com } Subject: LR White Question- Premature Polymerization. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Motorola SCG Chemical and Surface Analysis Lab is a multidiscipline analytical lab for wafer fab manufacturing in Phoenix, AZ. We are currently in need of a technician to provide TEM sample preparation support to our TEM operation. The candidate should have the following qualification. 1. Understanding of basic TEM imaging. 2. Experience or training in the following TEM sample preparation techniques: plane-view and cross-section TEM sample preparation using wedge polishing, dimpling; specific area cross-section with FIB. 3. Basic understanding of semiconductor devices and processing is highly desirable, but not required. 4. Applicants should have a A.A. degree in analytical technology, process good communication skill, and be able to work in a team-oriented environment.
Interested candidates should send resume directly to
Rebecca Ai MD P004 52nd St. Chemical and Surface Analysis Lab Semiconductor Component Group Motorola, Inc. 5005 E. McDowell Road Phoenix, AZ 85008 Ph. (602)-244-5775 Fax. (602)-244-6492 Email: RP3478-at-email.sps.mot.com
Bob: I have not found a listserver for AFM, but I have not looked for one for a while. You could check the AFM/Tunnelling section of the links on our site. There is a dozen good links, they may help anyway and perhaps point to the listserver - if one exists. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} Gentle folk, } } I believe that there has been mention of an atomic force microscopy } listserver at some time on this list. I am at home, recovering from surgery } and can't look through my filed mail at the office. } } We have begun doing some AFM work and I would like the address of } such a server, if it does exist. I'd like to make some good use of my } convalescence and learn more about the AFM before I return for work. } }
I have run into a couple of occassions of premature polumerization in the 10 or more years we have used LR White resin. In both cases the resin was old, ie over 1 year but was not associated with osmication. An possible explanation was suggested to me by Roy Gillett of London Resin a number of years ago who definitely recomended a shelf life of 12 months for catalysed resin.
Quote "The reason this pre-polymerisation occurs only with tissue must be something to do with a tissu constituent catalysing polymerisation. Older resin is much more susceptibe to this that fresh monomer becaue of the significant polymer growth that will inevitably have occurred in the monomer. The most likely 'endogenousd catalyst' from previous experience is likely to be an amine or peroxide moiety in the tissue"
We have had no problems since switching to buying uncatalysed resin and making up a new bottle as we run out of the old.
One point I have noticed in the discussions to date is some ambiguity between calalyst and accelerator. From my understanding the catalyst (benzoyl peroxide powder) must be added 24 hours before you start using a batch of resin and is necessary for both thermal (oven) and "cold" polymerization. The accelerator on the other hand is added to the final resin change for rapid "cold" polymerization without using an oven.
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
I have to embed some Drosophila melanogaster heads for transmission electron microscopy and prepare the same for scanning electron microscopy. The person requesting the work is interested in eye morphology. I have done a literature search and have come up with some relevant papers but my problem is that I will probably get these heads before I get the papers!! Working in the medical field, I have never embedded a sample with an exoskeleton before and am wondering if someone out there can give me an idea on the appropriate fixatives to use including fixation times and any little tricks I may need to know. Do the heads sink in the fixative or do I need to spin them down or use a vacuum? Is one resin better than another? How do they cut? Also, for SEM, which fixative do I use and for what duration? Again, are there any tricks to the dehydration and critical point drying procedures?
We have a TEM Siemens Elmiskop 101 in our lab. Recently, we encountered some problems with the high voltage controller. The cause was the bad connection of a K81A-type diode (electron tube-type diode) located in the power supply cabinet. We are looking for any K81A-type diode which could replace our old one.
Thank you for your help
Solvay Research and Technology Philippe Drouillon Rue de Ransbeek, 310 1120 Brussels (Belgium) Tel : (00 32) 2 264 24 47 Fax : (00 32) 2 264 20 55 Username : Philippe.Drouillon-at-solvay.com
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Dear colleagues,
Herewith I would like to thank, also on behalf of the other members of the "working group on accreditation of microscopical work" of the Dutch Society for Microscopy, all contributors and participants in the discussion on:
accreditation, callibration, standards, and ISO 9001.
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Image Analysis program for Nabisco in alignment with the Research Strateg= y. =
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and image analysis. Participates on project teams in both technical =
leadership and support roles depending on the needs of the project. Prov= ides =
proactive support to co-workers in the area of microscopy and image analy= sis =
in both identifying how the technology can be effectively used in project= s =
and providing user-friendly microscopy setups for co-workers. Has comman= d of =
food material science/physical chemistry which enables understanding and =
effective partnering in cross-functional teams/efforts.
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experience =B7 Degree in Food Material Science, Food Physical Chemistry, or related = field =B7 Develop a state-of-the-science microscopy & image analysis program fo= r =
Nabisco. In alignment with the Research Strategy, researches, identifies= , =
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company. If other instrumentation/ methodology is required that is not =
currently in-house, develops a database using outside sources =B7 Expertise in light (bright field, polarizing & fluorescence) and elec= tron =
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I've recently acquired a Oxford Instruments Link PCXA EDS X-Ray analyser - mid 1980s vintage but in good working order. My problem is the file format of the resulting spectra is markedly different to that from the contemporary QX2000/AN10000 analysers. I think its something to do with byte order being different for PC based processors. I need to be able to extract the files in order to label/print using ASCII readable spreadsheet software. If anyone has any experience/suggestions on the PCXA system, I'd be grateful to hear from them.
Thanks in advance,
Stu
****************************************************************** Stuart Kearns Electron Microbeam Laboratories Dept. of Geology tel: 44 (0)117 928 8204 University of Bristol fax: 44 (0)117 925 3385 Bristol BS8 1RJ UK email: Stuart.Kearns-at-bris.ac.uk ******************************************************************
Hello, my name is Lou Solebello and I am a research chemist/light microscopist with the JM Huber Company, Engineered Minerals Division in Macon Georgia. I am a newbie to SEM, and have not tried the Mquant software. A colleague of mine however, has been experiencing a lot of difficulty with getting the software to work properly. Does any one out there have experience and tips they would like to share? any help would be appreciated. I can be contacted by e-mail at either of the two addresses below.
First, for the TEM part: if you can get newly emerged adults, the cuticle will be soft and reasonably easy to section. By "newly emerged" I mean within a few hours. After the heads have assumed their normal shape (they inflate to rupture the pupal exoskeleton), but before they have tanned significantly. This assumes that the client isn't investigating something in the eye that changes subtly with tanning, or age after emergence.
The most difficult part of the eye for TEM are the crystalline cones. If the client isn't interested in them for TEM, dissect away the exoskeleton (cuticular area over the eye) and the underlaying c. cones. If possible, before fixation or embedding. (Waiting for the laughter to die here.) The easiest way to do this is chopping away with a glass knife after the blocks are ready.
You can handle the retinal tissues pretty much like any neural tissue. The client should already have the relevant fixation references, but routine fixation should be pretty normal. I used pH 7.2, 0.1-0.15 M buffers for freshwater crustaceans, standard Karnovsky's.
If s/he does want the cuticular and crystalline lenses ... wait for the references, or a response from a _Drosophila_ / small insect specialist--I worked on crustaceans (there are differences in the cuticle).
Use a *hard* resin with low viscosity. Other than that, I've seen no advantage for one type over another; except what works for you.
The heads should sink, if they don't, a *little* spinning won't hurt. By hand, I wouldn't use a centrifuge for this, even on the lowest speed. Mild vacuum would be better.
You mention "heads", so: is that what the client is bringing you, or are you planning on decapitating the critters? This would be good, if you can find a guillotine small enough. Best would be to then bisect the heads midsaggitally, if you can handle (orient, etc.) the resulting hemiheads. They may be pretty difficult to see to orient after OsO4, although the cuticle won't take up much osmium.
For SEM: treat like for TEM, except leaving the heads intact. CPD works well, and drying from HMDS can also (HMDS was originally used for Malphigian tubules in insects). Crystalline cones can be exposed nicely for SEM by dry-fracturing the heads after drying and mounting on the stubs. Read: hitting the eye with a razor blade, then gently blowing away the debris with a duster. (Also, look in through the back of an intact head.)
Mount on double-stickey *carbon conductive* tape, then run a thin line of silver paint to the head. Touch the wet paint to a sacrificial area of the head (one you don't care about), and draw a ring of Ag paint around the head, as close as you can get without touching it.
All of this last is to insure maximum conductivity of the specimen. The setae will charge like buggers otherwise, and you'll have bright little hairs, and lose structural details of both the setae and surrounding cuticle. Make certain that the heads have both excellent mechanical and electrical contact to the stub.
Phil } I have to embed some Drosophila melanogaster heads for transmission } electron microscopy and prepare the same for scanning electron } microscopy. The person requesting the work is interested in eye } morphology. I have done a literature search and have come up with some } relevant papers but my problem is that I will probably get these heads } before I get the papers!! Working in the medical field, I have never } embedded a sample with an exoskeleton before and am wondering if someone } out there can give me an idea on the appropriate fixatives to use } including fixation times and any little tricks I may need to know. Do } the heads sink in the fixative or do I need to spin them down or use a } vacuum? Is one resin better than another? How do they cut? Also, for } SEM, which fixative do I use and for what duration? Again, are there } any tricks to the dehydration and critical point drying procedures? } } Thanks in advance } } Sarah Ellis
I have to dry some bacteria grown on glass coverslips shortly for SEM from acetone to liquid CO2 i.e. critical point drying. I would appreciate advice in the next 24 hours about what times are recommended in the drier.
Unfortunately, the 'customer' has used quite a few 22 x 22 mm coverslips and I only have the original Polaron drier, without any specially-made coverslip holders, so it looks like three will fit in gauze baskets which I use for baby squid.
I spend my time embedding triatomino's legs. This is my method:
1)cut the heads 2)put 2-3 hs. in fixative medium ( I don't use transmition mic.) 3)deshidratation: ROH 70% (1*10 min) -- 80% ---- -- 90% ---- EtOH 100%(3*10 min) 4)EtOH 100% + Propilenoxide : (1:1) : ( 10-15 min) 5)Propilenoxide (10-15 min) 6)Propilenoxide + Durcupan (epoxi resine): (1:1) : ( 1 night) 8)open the recip. at 40 centigrade degrees ( 1h. 30 min) 9)put in new Durcupan at room temperature (1h.) 10)Put in new durcupan and orientation the head (1 night at 60 cent. degrees) Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
I am looking for any suggestions, advice with ongoing problem with weak immunostaining of semithin sections (0.5-1 micron) of low temperature Lowicryl K4M-embedded infiltrating ductal breast carcinoma.
After successful experiments on paraffin sections of the tissue, an incubation protocol for cytokeratin-8 antigen detection with immunoperoxidase-DAB procedure was applied to Lowicryl semithin sections, without the removal of the hydrophilic resin. The semithin sections were reacted with the antibody overnight at 4oC, and negative controls were performed by substituting the primary antibody with PBS. An increase of the staining contrast was obtained by posttreatment with OsO4. No staining was observed, as expected, at semithin sections used as controls. However, a very weak yellowish staining was observed at the sections treated with the which could not be evaluated. Trying to increase the intensity of the immunoreaction, I increased the concentration of the second Ab, the thickness of the sections (3-4 microns), and the incubation time of OsO4. The modifications took place at different experiments, but none of them was effective.
What should I do?? Any help would be greatly appreciated. Thank you in advance.
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Anyone know of a review article or bibliography on cathodoluminescence? I have one dating back to 1977 but I imagine there must be a more recent one. I did a lit search but came up negative. Any help would be appreciated. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
1. What is your experience using field emission SEM with EDXS.
2. What are the advantages and disadvantages. Is there sufficient beam current?
3. Would we be better off using tungsten filament source?, LAB6 source? for EDXS?
4. Imaging is also very important. As a reference point, we have a JEOL 6300F and are very pleased with the imaging at low keV. Are the current tungsten or LAB6 source microscopes being sold capable of providing the same level of image quality as a FE source microscope in the 1-5 keV range?
I really appreciate your time spent responding to these questions, however briefly.
There are many protocols that work. My experience indicates that 2% glut, 4% form in .1M PO4 works well for most TEM & SEM. A few minutes centrifugation at low speed (200rpm) hasn't damaged my samples. It helps to cut the proboscis off if you are not allowed to bisect the head. Standard EmBed resin has worked well for me--just allow infiltration overnight. Proper orientation can be the most difficult often requiring two or more embeddings, reorienting each time. SEM prep is very similar though I have found that critical point drying may require extended CO2 soaking times (30 to 60 minutes X 3). Some of our mutants have very little tissue in the head and are very susceptible to drying artifacts (they collapse). Some people leave the head attached to the body for ease of handling. I don't. Use low accelerating voltages in the SEM (10 kV or less) to minimize charging, etc. Good luck! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
} ... } 1. What is your experience using field emission SEM with EDXS.
... I shopped for a like instrument in '92 ...
} 2. What are the advantages and disadvantages. Is there sufficient } beam current?
Primarily your concerns for FE should be beam stability... enuf beam current was good question at the time ... but most FE guns were capable of delivering several nanonamps ... enuf for EDX.
}
} 3. Would we be better off using tungsten filament source?, LAB6 } source? for EDXS?
... better off, yes (... more stable ...), but you wouldn't get the low keV performance you mention below.
} 4. Imaging is also very important. As a reference point, we have a } JEOL 6300F and are very pleased with the imaging at low keV. Are the } current tungsten or LAB6 source microscopes being sold capable of } providing the same level of image quality as a FE source microscope in } the 1-5 keV range?
... you could come close with a LaB6 ... or a well designed W gun, but I still believe you'd find yourself optimizing the gun for low keV performance by varying the cathode-tip/wehnelt distance ... not something you want to be doing every day. Lastly, a FE gun is not likely going to provide you with the beam current stability you may want for EDX, especially for element mapping. I do remember there being methods of stabilizing a FE gun via feedback, but you want to make sure that the feedback isn't simply stabilizing the image contrast/brightness, but rather the beam current itself.
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} } 1. What is your experience using field emission SEM with EDXS.
3.5 years. AMRAY 1845FE w/Noran Voyager III, thin window. Plug and play. I work in an industrial services lab supporting many JIT manufacturing plants. Uptime is critical and the system has delivered as needed. The system has paid for itself over and over again. If I had the money, I'd upgrade all of our scopes to FE. } 2. What are the advantages and disadvantages...
Sufficient current has never been a problem. 18 nanoamps w/o apertures. We can easily swamp the detector with 100 micron aperture that provides a good balance between x-ray production and image quality. } 3. Would we be better off using tungsten filament source... We also have a highly modified AMRAY 1600 upgraded to a LaB6 last year. The scope is now more of a probe than an imaging tool. Light element EDS, 4 crystal WDS, CL, BSED, air-lock with sputtering gun, etc. Vinnie Casasanta, now at Charles Evans & Associates (Surface Sciences) built it specifically to support our materials development labs. Gun operation no problem. Just had to find the "sweet spot" in the saturation curve. Actually, Vinnie got enough current when he used an output valley rather than a peak because it provided a more stable beam. } } 4. Imaging is also very important...
I regularly use 500V to 1.5 keV for imaging at { { about 10k mag. At 5 keV, 50 thousand plus, no problem. With the LaB6 SEM, the instrument isn't tuned for imaging, it is just a directional current source so I can't really compare apples to apples.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
dear colleagues... a while ago, there was some chit-chat about a microscope from a company called Tasco...located in the northwest, I think... of course, now that i need the info i can't find it....I'd be grateful if someone could give me a lead to help me locate them... thank you... Ron Mervis ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Ronald F. Mervis, Ph.D. Chief Scientific Officer Neuro-Cognitive Research Laboratories Columbus, Ohio
} Lastly, a FE gun is not likely going to provide you with the beam } current stability you may want for EDX, especially for element mapping.
True, but I would imagine that any modern EDX system (ours included) provides some mechanism for normalizing maps for beam current variations. So don't worry about this one too much.
Regards, Rick Mott, PGT rick-at-pgt.com www.pgt.com
} remember there being methods of stabilizing a FE gun via feedback, but you wa } nt } to make sure that the feedback isn't simply stabilizing the image } contrast/brightness, but rather the beam current itself. }
I saw a paper a few years ago written jointly by someone from Hitachi and someone from Kevex, it described such a true feedback system, might be worth contacting either of those companies.
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
We agree with your assessment pertaining to the one year shelf life for catalysed LR White resin under ideal conditions. It is one of the reasons that we began shipping the uncatalysed resin several years ago. We will, if a researcher needs it, catalyse it and send it to them.
We also are in complete agreement on your last point about the catalyst and the accelerator.The catalyst is needed for both "hot and "cold" polymerisation, while the accelerator is added only for the rapid "cold" polymerization.
I have a Denton 502A vacuum evaporator that doesn't suck like it used to. I've tried looking at it and I'm getting pretty frustrated. The problem is it doesn't ROUGH Pump like it used to, it acts like it has a leak. Does anyone out there know of a company or individual who does service calls on Denton Vacuum evaporators that's on the west coast, preferrably in the San Francisco Bay Area? Please help me! I'm close to just hitting the thing with a hammer! Thanks for any information you might give me.
Frustrated in Berkeley,
Paula = ) psic-at-uclink4.berkeley.edu
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
First, do you *have* to CPD? I've used HMDS very successfully for bacteria, in some cases retaining the "slime" sheath (that depends more on dehydration). 5 min. changes, 100% Ac =} 1:1 Ac:HMDS =} 3 X 100% HMDS, dry at room temp or 60 C
Air drying straight from acetone can work for bacteria nicely--do you have time to experiment?
For CPD, use 3 to 5 changes of CO2, with 2 to 5 minutes soaking in CO2 between changes (time by slime coat). Your real problem is going to be turbulence on filling and emptying the chamber, so this will have to be done *very slowly* so has not to disturb the coverslips.
Note: this is Ed Basgal's design and idea!: A cover-slip holder can be quickly made by cutting notches in a polyethylene or glass tube (not tygon), just wider that the thickness of the coverslips, so the slips fit down into the notch:
_ |_ {--cover slip fitting into notch | | | |__|
place tube into a polyethylene syringe body or centrifuge tube that's been fenestrated by a mad perforator. Cap both ends.
Phil
} I have to dry some bacteria grown on glass coverslips shortly for SEM } from acetone to liquid CO2 i.e. critical point drying. I would appreciate } advice in the next 24 hours about what times are recommended in the } drier. } } Unfortunately, the 'customer' has used quite a few 22 x 22 mm } coverslips and I only have the original Polaron drier, without any } specially-made coverslip holders, so it looks like three will fit in gauze } baskets which I use for baby squid. } } Regards - Keith Ryan } Plymouth Marine Lab., UK
I spend my time embedding triatomino's legs. This is my method:
1)cut the heads 2)put 2-3 hs. in fixative medium ( I don't use transmition mic.) 3)deshidratation: ROH 70% (1*10 min) -- 80% ---- -- 90% ---- EtOH 100%(3*10 min) 4)EtOH 100% + Propilenoxide : (1:1) : ( 10-15 min) 5)Propilenoxide (10-15 min) 6)Propilenoxide + Durcupan (epoxi resine): (1:1) : ( 1 night) 8)open the recip. at 40 centigrade degrees ( 1h. 30 min) 9)put in new Durcupan at room temperature (1h.) 10)Put in new durcupan and orientation the head (1 night at 60 cent. degrees) Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
you posted a query re cathodoluminescence which I accidentally deleted, I have an expert here, if you want to mail me direct I'll pass on your query for review article(s)
Apologies if I've misspelled your name
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Subject; Charging for usage in the multiuser EM Unit
We are a university based multi-user EM Unit that is moving towards the 'real' world. Presently, all university users of our EM Unit are charged for the consumables they use (based on an honesty system) but charged for nothing else. Our bean counters are now requiring us to introduce equipment usage charges, labour charges and occupancy charges. At this point I have no idea how much they expect us to recover, the wording that is used is that those who use the service should contribute to the cost of running the service. Contribute is the key word.
I have no problem with this concept at all. My problem is however, how do we record the many 'chargables' and I am hoping that those of you out there who have introduced such charging systems to your Units and those who are working with such systems can help me.
Our EM Unit is multi-user Unit where 80% of the users come to the Unit and do the work themselves after we have trained them.
Equipment Charges Charging for electron microscope usage is simple enough taken just from a HT counter however how do you charge for the ultramicrotomes, microwave oven, tissue processor, pH meters, osmometer, enelargers, photo printer etc without having a lot of clipboards all over the place ?
Occupancy How do you keep records of all the users coming and goings from the Unit so that a sensible occupancy charge can be made ?
Labour Charges Easy to do by introducing a 'Job Card' system for the work we do.
All help and suggestions appreciated
Regards
Allan
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
I have examined microorganisms in the SEM directly, in their hydrated state with greatest success. I have used sputter coated or uncoated powdery mildew and rust colonies, and have about 30min before the spores dehydrate; any other treatment disturbes the delicate structures. The minute amounts of free water involved have not caused any problems with the vacuum or contamination; you obviously want to be sensible about this. One thing to remember, however, is that you will see the cells as they are, slime and all, which may obstruct interesting features of the bacteria.
If you have time to experiment, it might be well worth trying the simplest approach.
best wishes
Stephan Helfer
Sincerely +----------------------------------------------------------------- |Dr Stephan Helfer, SSO |Senior Mycologist - MSc Course Director | |Royal Botanic Garden Edinburgh, Inverleith Row, EDINBURGH EH3 5LR, |Scotland UK | |http://www.rbge.org.uk | |phone: +44 (0)131 552 7171 ext 280 | or +44 (0)131 459 0446-280 (direct digital VoiceMail line) |fax: +44 (0)131 552 0382 +------------------------------------------------------------------
Have you checked the air leak valve ? We once had a similar problem with our DV502 and it turned out to be a small leak in that valve. We have since put a filter right where the air goes in to prevent dust particles from collecting and causing poor seals.
I have a Denton 502A vacuum evaporator that doesn't suck like it used to. I've tried looking at it and I'm getting pretty frustrated. The problem is it doesn't ROUGH Pump like it used to, it acts like it has a leak. Does anyone out there know of a company or individual who does service calls on Denton Vacuum evaporators that's on the west coast, preferrably in the San Francisco Bay Area? Please help me! I'm close to just hitting the thing with a hammer! Thanks for any information you might give me.
Frustrated in Berkeley,
Paula = ) psic-at-uclink4.berkeley.edu
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Have a look at this book: B. G. Yacobi, D. B. Holt: Cathodoluminescence in inorganic solids. New York: Plenum 1990.
Dr. Hartmut S. Leipner Fachbereich Physik Friedemann-Bach-Platz 6 Martin-Luther-Universitat D-06108 Halle
Tel. +49-345-55 25 453 Web http://www.physik.uni-halle.de/Fachgruppen/Kristall/index.html
-----Original Message-----
Hello again, I have rec'd. a sample for TEM that may be infected by a herpes virus. I am wondering if there is a way of enhancing the tissue with any stains (tannic acid etc.) to optimally show the virus. The tissue has been fixed in 4%GA, cacodylate buffer. I will be processing today, so I appreciate any info asap. Sorry for the short notice. Thanks Linda Fox lfox1-at-wpo.it.luc.edu
A recent message contained a misprint. ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
------------------------------------------------------------------------=20 The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 =20 Hello All! =20 =20 I have a Denton 502A vacuum evaporator that doesn't suck like it used to=2E I've tried looking at it and I'm getting pretty frustrated=2E =20= The=20 problem is it doesn't ROUGH Pump like it used to, it acts like it has a=20 leak=2E Does anyone out there know of a company or individual who does service calls on Denton Vacuum evaporators that's on the west coast,=20 preferrably in the San Francisco Bay Area? Please help me! I'm close to just hitting the thing with a hammer!= =20 Thanks for any information you might give me=2E =20 =20 Frustrated in Berkeley, =20 =20 Paula =3D ) psic-at-uclink4=2Eberkeley=2Eedu =20 Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4=2Eberkeley=2Eedu =20 =20 =20 Paula; =20 Denton evaporators are almost indestructable, even hitting them with a= =20 hammer will not take them out of action for long=2E =20 The roughing and backing valves each have a bellows assembly inside=20 the valve housing, over time (several years) the bellows can develop=20 cracks resulting in leaks and long or impossible pump downs=2E This i= s=20 a likely possibility, but check the following first=2E =20 Do you have normal foreline pressure when both backing and roughing=20 valves are closed? This pressure should be 20mT or less=2E =20 =20 If this pressure is ok then what is the pressure when the backing=20 valve is opened? This too should be 20mT after a few minutes=2E =20 What is the best roughing pressure that you can achieve in 1, 5, and=20 10 minutes=2E =20 =20 If the foreline pressure and DP backing pressure are good but the=20 roughing pressure is poor, the main valve seal could be bad or the=20 seal around the bell jar could be poor=2E The bell jar could even have= =20 chips or hairline cracks=2E =20 =20 What is the best roughing pressure that you can get? If you can get=20 to high vacuum pumping what is the best pressure that you can get on=20 the penning gage? =20 Has the system ever been operated with both the backing and roughing=20 valves on at the same time, this is a bad thing! This will require=20 removal of the DP and inspection of the oil (color and amount)=2E =20 Usually when the backing and roughing valves have been opened=20 together, the DP stack will be displace upward resulting in poor high=20 vacuum performance or may even be impossible to hi vac pump=2E =20 Do you have manual roughing and backing valves (hand operated) or a=20 pneumatic system? Both of these use the bellows I described earlier=2E =20 If you have staff with vacuum experience you can probably fix this=20 yourself=2E Help is also available from Denton by contacting Jim Felc= o=20 at 609-439-9100=2E He may also be able to suggest a repair person in=20 your area=2E =20 Good Luck =20 =20 John Humenansky Braun Intertec Microscopy Department 6875 Washington Ave=2E So Minneapolis, MN 55439 612-942-4822
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Damian,
We have a Philips XL30 FEG w/ EDS and we get plenty of current (I have measured over 35 nA) that is very stable over time. In addition, we routinely image at 1-3 kV. All around it is a versatile instrument capable of excellent performance. I would say an FESEM is very capable of doing EDS.
On your comment about catalysts and accelerators, the JB-4 kit calls the benzol peroxide powder a catalyst and it is added to the infiltration solutions as well as the final solution. But, in the final mix, it is mixed in just prior to the solution B. JB-4 polymerization is done either at room temp or at 4 degrees centegrade. As a catalyst often accelerates a reaction, is there really a difference in calling it an accelerator or a catalyst? Just wondering.
Karen Pawlowski UT Dallas, UT Southwestern Medical Center
On Tue, 26 Aug 1997, IAN HALLETT wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have run into a couple of occassions of premature polumerization } in the 10 or more years we have used LR White resin. In both cases } the resin was old, ie over 1 year but was not associated with } osmication. An possible explanation was suggested to me by Roy } Gillett of London Resin a number of years ago who definitely } recomended a shelf life of 12 months for catalysed resin. } } Quote } "The reason this pre-polymerisation occurs only with tissue must be } something to do with a tissu constituent catalysing polymerisation. } Older resin is much more susceptibe to this that fresh monomer becaue } of the significant polymer growth that will inevitably have occurred } in the monomer. The most likely 'endogenousd catalyst' from } previous experience is likely to be an amine or peroxide moiety in } the tissue" } } We have had no problems since switching to buying uncatalysed resin } and making up a new bottle as we run out of the old. } } One point I have noticed in the discussions to date is some } ambiguity between calalyst and accelerator. From my understanding } the catalyst (benzoyl peroxide powder) must be added 24 hours before } you start using a batch of resin and is necessary for both thermal } (oven) and "cold" polymerization. The accelerator on the other hand } is added to the final resin change for rapid "cold" polymerization } without using an oven. } } Ian } } } } Ian Hallett } HortResearch } Mt Albert Research Centre } Private Bag 92 169 } Auckland, New Zealand } Fax 64-9-815 4201 } Telephone 64-9-849 3660 } EMail ihallett-at-hort.cri.nz }
This question has been around several times, so you might want to check the log of past postings.
We have run our facility as a fee-for-service lab for over 15 years now. Several things to keep in mind (debated hotly previously- so I hope I am not reopening the debate) is not to use University Subsidy to undercut your price to users outside your university.
Now, as to how do we determine fees for microtomes, etc. We know our service contract costs, we add depreciation to this and divide the total by the number of useable hours the instrument is available for use. This establishes our break-even cost. We add about 10% to this, since the instrument is down at times for service, etc.
Every piece of equipment has a sign-in/sign-out sheet. We send bills out once a month.
Consumables are charged at an average cost for a procedure. I.e. if you do a negative stain we know about what it costs for grids, stains, etc.
When someone in the laboratory does the work this cost is added to the charge.
Importantly, my consulting expertise comes free to University users. However, my time is charged for any outside work we do.
I would again urge you to consult past posts with regard to what is legal, ethical, and kind with regard to competition with outside suppliers of EM service who do not have the benefit of a University behind them. There are some fervently held beliefs on this score which should be heeded.
I hope this helps-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Wed, 27 Aug 1997, Richard Lander wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear all, Message on behalf of Allan Mitchell; } } Subject; Charging for usage in the multiuser EM Unit } } We are a university based multi-user EM Unit that is moving towards the } 'real' world. Presently, all university users of our EM Unit are charged } for the consumables they use (based on an honesty system) but charged for } nothing else. Our bean counters are now requiring us to introduce } equipment usage charges, labour charges and occupancy charges. At this } point I have no idea how much they expect us to recover, the wording that } is used is that those who use the service should contribute to the cost of } running the service. Contribute is the key word. } } I have no problem with this concept at all. My problem is however, how do } we record the many 'chargables' and I am hoping that those of you out there } who have introduced such charging systems to your Units and those who are } working with such systems can help me. } } Our EM Unit is multi-user Unit where 80% of the users come to the Unit and } do the work themselves after we have trained them. } } Equipment Charges } Charging for electron microscope usage is simple enough taken just from a } HT counter however how do you charge for the ultramicrotomes, microwave } oven, tissue processor, pH meters, osmometer, enelargers, photo printer etc } without having a lot of clipboards all over the place ? } } Occupancy } How do you keep records of all the users coming and goings from the Unit so } that a sensible occupancy charge can be made ? } } Labour Charges } Easy to do by introducing a 'Job Card' system for the work we do. } } All help and suggestions appreciated } } Regards } } Allan } } } } ----------------------------------------------------------------------- } Richard Lander } Electron Microscope Technician } South Campus Electron Microscope Unit } Otago School of Medical Sciences } P.O. Box 913 } Dunedin } New Zealand. } Tel. National 03 479 7301 Fax. National 03 479 7254 } } "Southernmost EM Unit in the World!" } ------------------------------------------------------------------------ } } }
In message {v02130500b028a30961ec-at-[128.32.175.193]} Paula Sicurello writes: } } I have a Denton 502A vacuum evaporator that doesn't suck like it } used to. I've tried looking at it and I'm getting pretty frustrated. The } problem is it doesn't ROUGH Pump like it used to, it acts like it has a } leak.
If it dopesn't rough pump, you probably DO have a leak or mechanical pump has a problem and it must be rather major and so perhaps easy to find. I have not maintained a Denton, but here are a few general things you can check, in case you havn't thought if these already.
1. Figure out ways to isolate parts of the system to check for leaks. To check if the leak is in the jar and its seal, or below the stage in the guts of the system, put a plug (eg. O-ringed fitted plate, like you have in your service kit for SEM or TEM) over the opening in the stage baseplate to the pumping system below and see if you can pump down.
2. If you can pump down, then the leak must be above the baseplate. If the bell jar has a rubber gasket mounted along the bottom edge, pull it back to check for chipping of the glass edge, or a crack. This can happen due to putting the jar onto the baseplate too hard and hitting a metal fitting on the stage. If so, clean out glass fragments, clean damaged edge with ethanol or acetone, dry with canned gas, fill with silicone bathtub caulk and gently reposition rubber seal. Then pump down just a little so that seal sets up under mild vacuum.
3. If system doesn't pump down as result of test in #1 above, then you have many more places to look!! : {( I suggest you check your mecnanical pump first. Can you pump down in the foreline area with the mechanical pump isolated from the rest of the system? If not, is the pump's oil level too low? Or if you havn't changed the mechanical pump's oil for a long time, try that for starters, as over time you can get moisture building up in the oil and that will certainly erode pump performance, usually at high vacuum end, tho. Do two changes, the first as a quick rinse - run on first oil change for about 5 minutes, then change it a second time.
4. Check all rubber or plastic hoses. First look at areas where tubing is clamped onto metal fittings, as hoses can eventually split there. Or tighten up hose clamps a little.
5. Valves and air inlets can also malfunction due to wear over years of use, or the lubrication drying out. You may need to open them up, clean and lubricate them.
Hope this helps and good luck.
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
A few years back, I contacted Denton about service and they had someone on the west coast who could do it. I believe it was Jim Falco, but you can contact Denton to find out for sure. At that time, I talked to Rob Specht (Marketing Manager) at (609)424-1012. Hope this helps.
Regards,
Bob *************************** Bob Citron Chiron Vision Claremont, CA USA (909)399-1311 Bob_Citron-at-cc.chiron.com ***************************
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Hello All!
I have a Denton 502A vacuum evaporator that doesn't suck like it used to. I've tried looking at it and I'm getting pretty frustrated. The problem is it doesn't ROUGH Pump like it used to, it acts like it has a leak. Does anyone out there know of a company or individual who does service calls on Denton Vacuum evaporators that's on the west coast, preferrably in the San Francisco Bay Area? Please help me! I'm close to just hitting the thing with a hammer! Thanks for any information you might give me.
Frustrated in Berkeley,
Paula = ) psic-at-uclink4.berkeley.edu
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I've gotten a couple of private emails on this. First, I didn't intend to imply that FE guns are necessarily less stable than LaB6 or W; this is not the case for many newer instruments, as several later postings on this topic have pointed out.
Second, a few people asked how you would correct an element map for current drift, regardless of gun type. One way is to collect a map of current per pixel corresponding to the element maps, by running a picoammeter signal through a voltage-to-frequency (V2F) converter. Most EDX systems have inputs for an external rate signal of this type. Scaling the X-ray counts using the current map counts at each pixel does the trick visually, although you're still left with pixel-to-pixel precision differences if you want to quantify the maps. Normalization can't fix that.
take a deep breath, relax, put down the hammer, pick up the phone and call Jim Falco, a service guru at Denton (609)439-9100. When I had a high vacuum leak in my unit, he walked me through step by step troubleshooting over the phone and I was able to find the problem (a microcrack in the flange in the main valve unit). He told me how much for the repair part and step by step how to install it, all of which worked out as he predicted.
I was very pleased with the assistance I received from Denton (I don't have any kind of service agreement or personal financial interest with them) and hope this type of after sales assistance is standard company policy that will continue in the future.
steve
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Dr. Steven Barlow, Associate Director EM Facility/Biology Department 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/EM_Facility
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Keith,
I too have used HMDS very successfully for a large number of bacteria species. HOWEVER, some bacteria require different protocols than others. Some species came through very well preserved with no evidence of shrinkage, others looked like they had been air dried from water! But with a little experimenting, you should find the right times. I used ethyl alcohol (I would think that acetone should be good as well): 25%, 50%, 70%, 80%, 95%, 100% x 3 15 min each, then a 3:1 EtOH:HMDS, 1:1, and 1:3 15 min ea, then HMDS 3X 15 min each, drain, then place in desicator to dry. Drying at 60deg ok too.
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Keith,
First, do you *have* to CPD? I've used HMDS very successfully for bacteria, in some cases retaining the "slime" sheath (that depends more on dehydration). 5 min. changes, 100% Ac =} 1:1 Ac:HMDS =} 3 X 100% HMDS, dry at room temp or 60 C
Air drying straight from acetone can work for bacteria nicely--do you have time to experiment?
For CPD, use 3 to 5 changes of CO2, with 2 to 5 minutes soaking in CO2 between changes (time by slime coat). Your real problem is going to be turbulence on filling and emptying the chamber, so this will have to be done *very slowly* so has not to disturb the coverslips.
Note: this is Ed Basgal's design and idea!: A cover-slip holder can be quickly made by cutting notches in a polyethylene or glass tube (not tygon), just wider that the thickness of the coverslips, so the slips fit down into the notch:
_ |_ {--cover slip fitting into notch | | | |__|
place tube into a polyethylene syringe body or centrifuge tube that's been fenestrated by a mad perforator. Cap both ends.
Phil
} I have to dry some bacteria grown on glass coverslips shortly for SEM } from acetone to liquid CO2 i.e. critical point drying. I would appreciate } advice in the next 24 hours about what times are recommended in the } drier. } } Unfortunately, the 'customer' has used quite a few 22 x 22 mm } coverslips and I only have the original Polaron drier, without any } specially-made coverslip holders, so it looks like three will fit in gauze } baskets which I use for baby squid. } } Regards - Keith Ryan } Plymouth Marine Lab., UK
(IMA Internet Exchange 2.1 Enterprise) id 00194947; Tue, 26 Aug 97 19:56:17 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ns2.baxter.com (8.8.0/8.8.0) with SMTP id UAA24598; Tue, 26 Aug 1997 20:03:38 -0500 (CDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA08994 for dist-Microscopy; Tue, 26 Aug 1997 17:16:31 -0500 Received: from ux1.cso.uiuc.edu (ux1.cso.uiuc.edu [128.174.5.59]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA08991 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 26 Aug 1997 17:16:30 -0500 Received: from [130.126.25.7] (florence-8.slip.uiuc.edu [130.126.26.132]) by ux1.cso.uiuc.edu (8.8.5/8.8.5) with SMTP id RAA07717; Tue, 26 Aug 1997 17:23:45 -0500 (CDT) X-Sender: oshel-at-staff.uiuc.edu Message-Id: {v02120d01b0290a6b20d7-at-[130.126.25.7]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
On Wed, 27 Aug 1997, Linda Fox wrote:
} Date: Wed, 27 Aug 1997 07:33:38 -0500 } From: Linda Fox {lfox1-at-wpo.it.luc.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: enhancing herpes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello again, } I have rec'd. a sample for TEM that may be infected by a herpes } virus. I am wondering if there is a way of enhancing the tissue with } any stains (tannic acid etc.) to optimally show the virus. The tissue } has been fixed in 4%GA, cacodylate buffer. I will be processing } today, so I appreciate any info asap. Sorry for the short notice. } Thanks } Linda Fox lfox1-at-wpo.it.luc.edu
Unless you're going to do immunoEM, any fixes using glut, Os, and UA will be fine. Infected cells should have 100 nm nucleocapsids in the nucleus and 200 +/- nm complete virions in the cytoplasm. Nucleocapsids may bud from nuclear, intracytoplasmic, and plasma membranes. You can cut thick sectins and stain them with toluidine blue, Paragon, multiple stain, etc. and look for abnormal areas before selecting areas for thins. Contact me directly if you have questions.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I remember there being some mention in the past of techniques to bring a computer display into harmony with the a printout regarding color hues, intensity, etc. Does anyone remember this or have any suggestions to add? I seem to recall there being certain software tricks or packages involved.
For background, we just purchased an Epson Photo Stylus color printer and a Polaroid Sprintscan 35 with PathScan Enabler, which seem to do a great job for scanning and printing light microscope slides to get an overview of the whole section. The only problem is that the colors on the printout don't exactly match the screen display (computer running Windows 95), and neither are perfect in comparison to seeing the slide under the microscope. We could live with what we get on the screen if only the printouts would match. A colleague has been playing with settings in Adobe Photoshop (CMYK vs. RGB) and with printer ICM (??) setting, but they only seem to make things worse.
Oh and I should mention we haven't yet contacted the vendors to get their input so perhaps they will have a simple suggestion. But in the mean time any help from the list would be appreciated!
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
I am looking for a US supplier for Nuclepore gridded filters. I would also like the phone number or web site for the Nuclepore Corporation. Please send the information to my email address: gondo-at-sprynet.com.
Analytical Consumer, a monthly newsletter for analytical laboratories, is doing a survey of users of all types of scanning probe microscopes. Each month, we survey users of one kind of analytical equipment, asking what they use, why they bought it, and what they think of its performance and support. Then we report what the labs say. We are not associated with any organization or instrument company, and we accept no advertising.
If you use SPM and would be willing to be part of the survey, send me an e-mail (address below), and I'll send you the short list of questions. Every one who replies receives a copy of the final report, of course.
Thanks, Jo Rita
Jo Rita Jordan, PhD Editor and Publisher Analytical Consumer
There were some questions a while back on the list about the file format used by the Oxford/Link ISIS .
I'm currently using the export to EMMFF to get the data out of the link system, but it would still be useful to have info on the format for batch conversions of large data sets. ( Or for when you don't have access to the ISIS software. )
If anyone has any more authoritive info, I'ld like to hear it. In the meantime, from reverse engineering some of our data files and comparing them with the EMMFF output, I have: ( Note: I don't know what variable info may be in the header that might be different in a different setup -- this is from a small sample of files. )
Partial Format of .SPE files: ( offsets are zero based )
Spectrum is the last 1024 ints of the file.
CHOFFSET is float at word [384] RealTime is int at word [130] LiveTime is int at word [128] ( or are these perhaps the low order of a double-word long int ? ) RealTime and LiveTime are both in microsecs.
A string containing title and analyst starts at byte[10] ( don't know if this is fixed or variable length, and whether it's split into two separate fields. )
This may be enough for a partial conversion. If anyone has any more info to add, please let me know.
[ BTW: I've found Python {http://www.python.org/} to be a good tool for this sort of reverse-engineering/investigative-programming. The array classes even have byteswap() methods, which, since I'm doing this conversion on a Mac, and ISIS files come from a PC, come in quite handy. I also have a new version of a DTSA -} EMMFF batch converted written in Python, if anyone is interested. ]
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |--- All power corrupts and obsolete power corrupts obsoletely." - Ted Nelson
} Date: Wed, 27 Aug 1997 09:13:28 -0500 (CDT) } From: {kna101-at-utdallas.edu} } To: IAN HALLETT {ihallett-at-hort.cri.nz} } Cc: microscopy-at-sparc5.microscopy.com } Subject: RE: LR White Question- Premature Polymerization.
} Ian, } } On your comment about catalysts and accelerators, the JB-4 kit } calls the benzol peroxide powder a catalyst and it is added to the } infiltration solutions as well as the final solution. But, in the final } mix, it is mixed in just prior to the solution B. JB-4 polymerization is } done either at room temp or at 4 degrees centegrade. As a catalyst often } accelerates a reaction, is there really a difference in calling it an } accelerator or a catalyst? Just wondering. } } Karen Pawlowski } UT Dallas, UT Southwestern Medical Center } Karen
In the case of LR White resin there is a difference in that the catalyst powder must be added to the resin well before it is used (at least 24h) and is used to "activate" the bottle of resin to allow polymerisation. The shelf life of the resin starts from this time. Initially all LR White resin was sold catalysed which caused some problems with old stock.
The accelerator is added only when a "cold" cure is requried - I'm not sure what it is (comes as a liquid), we never use it. The "cold" cure is a strong exothermic reaction hence the "" around cold.
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
John J. Bozzola wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Anyone know of a review article or bibliography on cathodoluminescence? I } have one dating back to 1977 but I imagine there must be a more recent one. } I did a lit search but came up negative. Any help would be appreciated. } Thanks. } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901-4402 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
A quick follow-up to John's question re: cathodluminescence bibio..
There was a device, sold in the UK, which did cathodluminescence on a light microscope. Does anyone have lit, info, source of supply for US?
At 01:05 PM 8/26/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The disadvantages are: With cold field emission the current fluctuations from the gun have to be compensated when you are doing quantitative EDS
We find we dont do any quant with our 4500 (we also have a Cameca Probe) so that is no concern
Cold field has the lowest beam current of any emitter. But it is still enough to saturate the EDS detector and also for our cathodoluminescence system
Information to the contrary from sellers of Hot field emitter guns is too pessimistic
} 3. Would we be better off using tungsten filament source?, LAB6 } source? for EDXS? } In practice, No.
} 4. Imaging is also very important. As a reference point, we have a } JEOL 6300F and are very pleased with the imaging at low keV. Are the } current tungsten or LAB6 source microscopes being sold capable of } providing the same level of image quality as a FE source microscope in } the 1-5 keV range? }
Absolutely NOT!. We had a S-900 FESEM and bought the 4500 so we could apply the brilliantly improved resolution and low kV we were getting to large specimens. It as forever changed how we do our microscopy.
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Those who have tried to reach the Web site for next year's International Congress on Electron Microscopy to be held in Cancun Mexico may have been disappointed in the last couple of days. I have just called Mexico. The Web site is being updated. It is expected to up again early next week at
http://icem.inin.mx
Plan on attending the meeting! .
** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
I understand that information on the next International Stereology Course was posted earlier on the listserver. I have just subscribed and would be very grateful if someone could forward me the posting or any relevant information.
Many Thanks Teeba Lundy lundyt-at-agresearch.cri.nz Wallaceville Animal Research Centre Upper Hutt NEW ZEALAND
anyone out there who has a philips 400 tem that could give me simple instructions for alignment and filament saturation? i'm new in a lab that has no one (tech retired and just left) that is familiar with this instrument. i've been doing okay so far but it would be nice to hear from someone who has this instrument.
It is quite impossible to charge for each grid, piece of glass etc. For many years I used a scheme to just charge for EM time, sheet film and prints. The prints were charged by weighing customer weighing the current paper box before and after and recording this nearest gram in a logbook. Weighing the box doubled the returns on the previous "I used so many sheets" - people are honest when you can verify their honesty. Paper paid for developer and fixer and about 15% extra covering 35mm TEM film too. SEM 120 roll film was charged for, but a 35mm camera with 50 ASA film and a half decent macro lens is just as good on the SEM and can be fully automatically integrated.
The SEM/ TEM charges had to pay for all other consumable in the lab. Filament time not kv should be used because for most situations, especially high res its better to leave the kV on. For the probe its best to charge for time booked, because the filament too needs to remain on.
Its far more efficient and cost effective to purchase centrally. If every user needs to buy grids, osmium and buffer etc. an awful waste occurs (which should be good for my business). Try to teach users to use small volumes. One problem are lab users who just prepare their samples in your lab and look at them elsewhere - or just use LM. Overall this system worked well for many years.
I charged $12 for EM hour and received a maintenance grant of $12000 which was cut to $5000 after some years and then a new, half-baked VC (CEO/ or president) declared: Its user pays; no maintenance. That is when the arguments get interesting. Note a poor CEO makes up policy and is not interested to make policy work in a real world.
That lab had a considerable teaching function for undergrads, but also had a hundred grad students a year. If they spent 20% of their time in the EMU on related activities, the Unit should have been top ranking in box top counts. The 3 EM busy unit was run with one and a half staff only. That is a bit of a tricky thing to handle. As a result, in terms of user-pays would have excelled 'in the real world'. Admin, library and computing took half of all Gov fees, about$20000 annually for the students. The departments received the other half and the departments returned generally $1000 to PhD students.
Do the arithmetic any way and it would be obvious that the EMU more than paid its way. What that VC wanted was to count every $ twice. User pays was a scheme to get more money from nothing, which is a poor economic principle.
The EM Committee voted against increasing hourly fees. To absorb the lost maintenance would have meant at least a doubling of hourly fees. Higher fees result in less usage. Eventually the VC wanted to charge for labour too. That would have required charges of about $70/filament hour.
Get me right, I am not against 'user pays'. It could have worked if the students received say $5000 of there funding to spent on campus based research. In my experience the phrase 'User pays' is used to give legitimacy to fancy schemes, like charging effectively twice, increasing resources for admin. and decreasing the dollar going to the universities primary functions: Learning and research.
That VC, after twelve long years was shown the door - but he still received a 'golden hand shake'. His legacy is a smallish uni with a debt of $29 million and a lot of run down facilities and a very demoralised staff.
An administrations role is to facilitate learning and research. Sure, the place has too operate within budget, but beware of anything that makes administration more complex. Charging and other admin tasks may be necessary, but the effort detracts from research and to an extend is counter-productive. Charging must make a lab less efficient, so use the simplest system that can be devised.
Beware of administrators spouting half-smart slogans. Charging is meant to extract more money from somewhere, but nobody has put more money into the system. So it is just a way of re-allocating resources, usually do the detriment of the university's primary functions. Cheers, its no good crying about that. Jim Darley
This is really rather out of topic, but would any of you by any chance know of an existing listserver (just like Microscopy) that discusses ischemia, reperfusion injury, animal and cellular models and related topics?
Hope you could help.
Thanks!
Ginny
------------------ GINNY E. CRUZ Section of Immunopathogenesis, Institute of Immunological Science Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 e-mail: gcruz-at-imm.hokudai.ac.jp ------------------
} Dear All of the Microscopy Net: } } This is really rather out of topic, but would any of you } by any chance know of an existing listserver (just like } Microscopy) that discusses ischemia, reperfusion injury, } animal and cellular models and related topics? } } Hope you could help. } } Thanks! } } Ginny } } } ------------------ } GINNY E. CRUZ } Section of Immunopathogenesis, Institute of Immunological Science } Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN } Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 } e-mail: gcruz-at-imm.hokudai.ac.jp } ------------------
------------------ GINNY E. CRUZ Section of Immunopathogenesis, Institute of Immunological Science Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 e-mail: gcruz-at-imm.hokudai.ac.jp ------------------
Hello all ! I have a problem in determining the divergence angle of our TEM Jeol 2010=20 for the different modes of this microscope. Indeed, three divergence angles= =20 are available for imaging studies called : alpha=3D1, 2, 3, which should be= =20 used from high magnification values (} mag 400kX) to low maginifcations=20 ( {100 kX) and then should correspond to increasing divergence angle from 1= =20 to 3. I have tried to measure this angle, by classical method (on E.D=20 patterns obtained with a condensed electron beam), but obtained values were= =20 absurd, in total contradiction with expected ones. Jeol company from France= =20 cannot give me any explanation for this, nor another measurement procedure.= =20 Can anyone help me ? Thanks Bernadette Domenges CRISMAT-ISMRA Boulevard du Marechal Juin 14050 CAEN CEDEX - FRANCE E-mail : domenges-at-crismat.ismra.fr Fax : (33) 02 31 95 16 00 T=E9l : (33) 02 31 45 26 32
DuPont Medical Products Biotechnology Systems Division 31 Pecks Lane Newtown, CT 06470-5509
Tel: (800) 551-2121 Fax: (203) 270-2166
http://www.sorvall.com
They don't distribute through other suppliers or distributors.
Regards,
Rick Powell
****************************************************************** * PLEASE NOTE MY NEW E-MAIL ADDRESS: rpowell-at-mail.lihti.org * * NANOPROBES GENERAL E-MAIL ADDRESS: nano-at-mail.lihti.org * * * * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
} Hi All, } } I need to find a supplier for the Sorvall GLC-2, general } laboratory centrifuge. We inherited one and need some more test tube } holders. Thanks. } } Karen Pawlowski } UT Dallas/UTSW Medical Center
} } } } Dear All of the Microscopy Net: } } } } This is really rather out of topic, but would any of you } } by any chance know of an existing listserver (just like } } Microscopy) that discusses ischemia, reperfusion injury, } } animal and cellular models and related topics? } } } } Hope you could help. } } } } Thanks! } } } } Ginny } } } } } } ------------------ } } GINNY E. CRUZ } } Section of Immunopathogenesis, Institute of Immunological Science } } Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN } } Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 } } e-mail: gcruz-at-imm.hokudai.ac.jp } } ------------------ } } ------------------ } GINNY E. CRUZ } Section of Immunopathogenesis, Institute of Immunological Science } Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN } Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 } e-mail: gcruz-at-imm.hokudai.ac.jp } ------------------
------------------ GINNY E. CRUZ Section of Immunopathogenesis, Institute of Immunological Science Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 e-mail: gcruz-at-imm.hokudai.ac.jp ------------------
Some people mentioned that a premature polymerisation of the resin might be avoided if the catalyst is added to the resin just prior to the embedding.
I looked into several catalogues from different vendors but only found kits where the resin (presumably containing the catalyst) and the accelerator were offered.
I would like to ask for inputs which company sells LR White with the catalyst seperate (preferably in Europe).
Hans-Martin ************************************************************** Hans-Martin Vaihinger Ruhr-University of Bochum Comparative Endocrinology Research Section Building ND 5/37 44780 Bochum GERMANY ********************************************************* phone ++49 234 700 4329 fax ++49 234 709 4551 e-mail Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de
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Hello-
I am looking for a US supplier for Nuclepore gridded filters. I would also like the phone number or web site for the Nuclepore Corporation. Please send the information to my email address: gondo-at-sprynet.com.
I buy the LR White from Ted Pella Inc. and it comes separate.
Bob
On Thu, 28 Aug 1997 Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } To the people discussing LR White problems } } Some people mentioned that a premature polymerisation of the resin } might be avoided if the catalyst is added to the resin just prior to } the embedding. } } I looked into several catalogues from different vendors but only } found kits where the resin (presumably containing the catalyst) and } the accelerator were offered. } } I would like to ask for inputs which company sells LR White with the } catalyst seperate (preferably in Europe). } } Hans-Martin } ************************************************************** } Hans-Martin Vaihinger } Ruhr-University of Bochum } Comparative Endocrinology Research Section } Building ND 5/37 } 44780 Bochum } GERMANY } ********************************************************* } phone ++49 234 700 4329 } fax ++49 234 709 4551 } e-mail Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de }
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I am trying to find Roger Waldock. I recall that he sold EDM (spark machining) instruments for slicing and coring of samples for metallography, TEM,...etc. sample preparation. The instrument that we are interested in buying is the Servomet. I believe that it is/was produced in Cambridge, England. If anyone can help in locating Roger and or who sells these in the USA I would appreciate it.
Thanks, Mark A. Wall L-350 Chem. & Mat. Sci. Dept. Lawrence Livermore National Lab 7000 East Ave. Livermore, CA 94550 USA ph. 510 423-7162 fx. 510 422-6892
} The instrument that we are interested in buying is the } Servomet. I believe that it is/was produced in Cambridge, England. If anyone } can help in locating Roger and or who sells these in the USA I would } appreciate it. } I do, however, have some copies (circa 1990) of EDM Digest, more recent issues of American Tool, Die & Stamping News and Metal Forming. I'd be happy to send you any or all of these if you could use them. Yours, Bill Tivol
My SEM books are quite vague regarding what channeling contrast on the SEM is. Can anyone out there tell me 1) What is the mechanism of channeling contrast on the SEM? 2) What detectors/equipment is necessary to do channeling contrast on the SEM? 3) What are the sample and sample prep. requirements for channeling contrast on the SEM?
DuPont Medical Products Biotechnology Systems Division 31 Pecks Lane Newtown, CT 06470-5509
Tel: (800) 551-2121 Fax: (203) 270-2166
http://www.sorvall.com
They don't distribute through other suppliers or distributors.
Regards,
Rick Powell
****************************************************************** * PLEASE NOTE MY NEW E-MAIL ADDRESS: rpowell-at-mail.lihti.org * * NANOPROBES GENERAL E-MAIL ADDRESS: nano-at-mail.lihti.org * * * * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
} Hi All, } } I need to find a supplier for the Sorvall GLC-2, general } laboratory centrifuge. We inherited one and need some more test tube } holders. Thanks. } } Karen Pawlowski } UT Dallas/UTSW Medical Center
I can't help you much with locating Roger, but I thought I would make you=
aware that we do offer a small Spark Erosion Unit for TEM applications. = It is an inexpensive unit and is designed for small samples. You can get so= me limited information on our web site or I would be pleased to send you a complete data sheet on it.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com =
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
Message text written by "Mark Wall" } I am trying to find Roger Waldock. I recall that he sold EDM (spark machining) instruments for slicing and coring of samples for metallography, TEM,...etc. sample preparation. The instrument that we are interested in buying is th= e Servomet. I believe that it is/was produced in Cambridge, England. If anyone can help in locating Roger and or who sells these in the USA I would appreciate it.
Every now and again, with semi-thin sections made from Epon-araldite plastic (0.5 microns thick) of nerve or muscle, I get a wrinkled result that looks as though the entire section has fractured. I've tried adjusting the temperature of the hot plate, or even giving it only 30 seconds on the hot plate, I've tried cutting wide rather than long, I've tried trimming away all the excess plastic around the tissue area, and I've tried cutting thinner sections, but in all cases, once I get this "cracked" appearance to the section, I have great difficulty cutting it to give me secitions that don't have these networked wrinkles.
Does anyone have any thoughts as to how I might avoid this problem, before I go insane!!
Garry, It has been suggested to me to try using 40% acetone/dH2O instead of dH2O when semi-thin sectioning. I tried it with just Spurr's and it worked pretty good (but this was a botanical sample). Good Luck! Tracey Pepper Iowa State Univ.
Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } To the people discussing LR White problems } } Some people mentioned that a premature polymerisation of the resin } might be avoided if the catalyst is added to the resin just prior to } the embedding. } } I looked into several catalogues from different vendors but only } found kits where the resin (presumably containing the catalyst) and } the accelerator were offered. } } I would like to ask for inputs which company sells LR White with the } catalyst seperate (preferably in Europe). } } Hans-Martin } ************************************************************** } Hans-Martin Vaihinger } Ruhr-University of Bochum } Comparative Endocrinology Research Section } Building ND 5/37 } 44780 Bochum } GERMANY } ********************************************************* } phone ++49 234 700 4329 } fax ++49 234 709 4551 } e-mail Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de
We are on such company and we sell direct to Germany or we can give you the names of one of our agents in Europe if you would like to buy from them. Please let me know.
Currently we have a JEOL JAMP 30 auger microprobe with Link AN 10/55S light element detector for anyone who is interested. Please email me at this address and indicate "microprobe" in your subject header or fax an information request to M.W. Rigler at MAS, Inc. 770-368-8256. The fax will ge a faster response.
Cheri Moss asked for a reference on channeling contrast in SEM. I suggest trying Chapter 3 in the book 'Advanced Scanning Electron Microscopy & X-Ray Microanalysis', by Dale Newbury, et. al., Plenum Press, 1986,ISBN 0-306-42140-2.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Regarding the necessity of charging for EM services, it is now officially near the end of the silly season, so I thought I would like to share with you all this little story. I hope you find it amusing!
I will go back a few hundred years to the Middle East, to visit a man called Nasr-ed-Din. He was a local official who worked as a town clerk, magistrate, and letter writer, since he was the only learned man in his town or village. A man in his position would have been called a Mullah in Persian, and in Turkish a HODJA, which is how I will call him from now on.
The Hodja had heard about some Sufis who claimed that they could achieve enlightenment by going for a long time without food, and he thought that this sounded like a good idea. But it also seemed a bit dangerous, so he thought he would try it on his donkey first. He normally gave the animal 20 scoops of grain a day, so for the next week he gave it 19, then the week after 18, and so on. By the time he had reached 10, the donkey was beginning to get weak and to wander about as if in a dream.
"Excellent" thought the Hodja, "it is beginning to learn the art of meditation!"
But just before he was getting down to 3 scoops a day, the donkey died.
"Inconsiderate beast" he cried. "Dropping dead before the experiment was complete. If only it had lived a few weeks longer, it might have achieved full enlightenment!"
All over the world, in university science departments, numbers have been falling. Typically, since around 1970;
- Student numbers have dropped to about 75% of their original number;
- Teaching staff to about 50%;
- Technical staff to about 25%.
This last figure has put an enormous burden on the research as well as the teaching effort. Whenever A retires, the government body says "You don't need a replacement - can't B and C share his job? And so on ... it is predicted that by the year 2010 such a department will be running on no staff at all! But it must expire before then, once it gets below a critical (m)ass.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
We handle a fair number of muscle, and nerve biopsies and are usually able to avoid wrinkles by flattening with a heat pen while the section is still floating in the boat. Although, every so often we do have a specimen that has such internal elasticity (tension) that occasional wrinkles do occur. These are not apparent in the ultra-thin section (also flattened with a heat pen). You may want to check the specimen protocols if this is a consistent problem. Ensure that the specimen is clamped or tied with the correct amount of tension when accepted from surgery and during fixation.
Peter O. Steele, PhD All Children's Hospital St. Petersburg, FL
} } } Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 08/28/97 02:25pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Every now and again, with semi-thin sections made from Epon-araldite plastic (0.5 microns thick) of nerve or muscle, I get a wrinkled result that looks as though the entire section has fractured. I've tried adjusting the temperature of the hot plate, or even giving it only 30 seconds on the hot plate, I've tried cutting wide rather than long, I've tried trimming away all the excess plastic around the tissue area, and I've tried cutting thinner sections, but in all cases, once I get this "cracked" appearance to the section, I have great difficulty cutting it to give me secitions that don't have these networked wrinkles.
Does anyone have any thoughts as to how I might avoid this problem, before I go insane!!
Dear Sirs: I am working on the sputtering NiFe/Mo magnetic multilayers by using HREM. The interface between the NiFe and Mo layers may be a kind of NiFeMo alloy, but I can't make decision about the idea. So I hope to know the NiFeMo phase diagram and the solution degree between NiFe and Mo.
I am looking for the help from you and please tell me the references where I can find the NiFeMo phase diagram.
The Department of Biological Sciences of the Exacts and Natural Sciences Faculty of the Buenos Aires University are looking for donation of a TEM and-or SEM equipment, not older than 15 years and still woorking. We will paid any handling and shipping cost. It would be very helpfull for us any other kind of assistance. Thank you vey much in advance.
Please respond to me at the addresses and numbers listed below.
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA
Z.Zhang's Postgraduate students wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Sirs: } I am working on the sputtering NiFe/Mo magnetic multilayers by } using HREM. The interface between the NiFe and Mo layers may be a kind } of NiFeMo alloy, but I can't make decision about the idea. So I hope to } know the NiFeMo phase diagram and the solution degree between NiFe and } Mo. } } I am looking for the help from you and please tell me the } references where I can find the NiFeMo phase diagram. } } Yours Sincerely } } Gao Yihua
Gao Yihua,
Try with ASM book of Phase Diagrams. More info about this book you may found at: http://http://www.ASM-INTL.ORG. They have single phase diagram delivery service.
"Frustration" is a kind word for it. I fought for 2 whole years with a project requiring embedding of skin (muscle and epithelium). It was one of the worst fights I ever conducted in my association with LM - TEM. I finally was able to solve it and wrote a paper - Crowley, Hildegard, H., Elimination or Reduction of Wrinkles in Semithin Epoxy Sections.....1988.Stain Technology, Vol 64, No. 5, p 221. From your short description I surmise that you need to change your protocol, starting with dehydration (perhaps earlier). You need to change your embedding medium. Araldite-Epon-DDSA is "too soft" or not crosslinked enough for your application. The differences in viscosities between Araldite and Epon is so great that muscle tissue may act as a diffusion barrier to the Araldite, thus allowing the seperation of monomers before polymerization. I personally would abondon that formulation for that purpose, but if there were reason not to, I would take other measures. Please call me if you cannot get the paper or have further questions. I understand your frustration completely! I cannot go into a very lengthy discussion at this time, but I suggest for a trial the following: Take some of your wrinkled sections, soak the slides in water for 30 minutes or so, dry off the slides with tissue paper. Place slides in a rack. Outfit a vaccum jar with Na pentoxide ( a flat dish of some sort - use a generous amount). Put the dish in the bottom of the jar and cover the dish with a piece of large filter paper. Put the inset of the vaccum jar back in place. Set the rack of slides on the inset (rack of slides will be elevated over the pentoxide). Grease the rim of the lid well and close. Gradually pull a good vaccum on the jar. Leave the slides for 48 hours. Gradually admit air to the jar. Inspect. It may not work, but it is worth a try. If it works, you are in clover. If not, call me. 303-871-3026. Do not get discouraged. You CAN solve this problem. Bye, Hildy Crowley University of Denver
What do routine labs do with uranyl acetate waste. Ours has been picked up by in house safety department and is being stored in drums as there isn't a company that will pick up to discard. Is anyone else having this problem. It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with copious amounts of sterile distilled water. Any imput will be appreciated. Many thanxs
} My SEM books are quite vague regarding what channeling contrast on the } SEM is. Can anyone out there tell me } 1) What is the mechanism of channeling contrast on the SEM? Basically, the BSE yield is dependant on the relative beam/crystal orientation for a sample, with ``large'' changes in BSE yield occourring over small changes in angle near the Bragg conditions for the various crystal planes in the sample.
} 2) What detectors/equipment is necessary to do channeling contrast on } the SEM? Channeling contrast generally is on the order of only a few percent, so maximization of signal is important. High BSE collection efficiencies are desirable, as well as a large probe current. Since the signal yield is a function of the electron trajectory relative to the crystal, a small convergence angle for the scanning probe is also a good idea.
} 3) What are the sample and sample prep. requirements for channeling } contrast on the SEM? -A ``smooth'' surface, since the ECC signal is easily swamped by topographic contrast. Likewise, a sample of farily uniform composition is also helpful beceause of Z contrast. -A clean, undisturbed crystal surface. This is beceause ECC requires a coherent electron beam interacting with the crystal, and coherency is lost over a fairly short distance of travel through any surface layers. For both these reasons, I prepare most of my channeling contrast samples by electropolishing.
Note: if channeling contrast is being used for grain boundries and the like, the contrast can often be picked up by a secondary or E-T style detector beceause of the linkage between BSE yield and secondary yield. A further note: ECC for imaging crystal defects follows all the points above, with the addition that the sample must be oriented relative to the beam in a fashion strongly analogous to orienting a TEM sample for diffraction contrast. This usually requires either large crystals or the ability on your microscope to do selected area channeling, so that the desired channeling condition can be established. } Have fun with ECC! Ben Simkin, simkin-at-egr.msu.edu Michigan State University dept. Materials Science
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Dear Sirs: I am working on the sputtering NiFe/Mo magnetic multilayers by using HREM. The interface between the NiFe and Mo layers may be a kind of NiFeMo alloy, but I can't make decision about the idea. So I hope to know the NiFeMo phase diagram and the solution degree between NiFe and Mo.
I am looking for the help from you and please tell me the references where I can find the NiFeMo phase diagram.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sirs: I am working on the sputtering NiFe/Mo magnetic multilayers by using HREM. The interface between the NiFe and Mo layers may be a kind of NiFeMo alloy, but I can't make decision about the idea. So I hope to know the NiFeMo phase diagram and the solution degree between NiFe and Mo.
I am looking for the help from you and please tell me the references where I can find the NiFeMo phase diagram.
Hello all, I need to look at the surface morphology of some yeast cells and as I'm a physics sort of person I need a little help.
I going to be using an ESEM, but what is the best way to prepare yeast cells for this sort of examination?
A step by step answer or reference would be great.
many thanks
David
Dr. David C. Bell Room 13-1018 E-Mail: dcb-at-MIT.EDU Center for Mat. Sci. and Eng. PH: (617) 253-3317 Massachusetts Institute of Technology FAX: (617) 258-6478 77 Massachusetts Ave, Cambridge, MA 02139-4307
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