we are using the PIPS for several years already and I have to admit that we cleaned the inner surface of the work chamber only in *very* long intervals. This is only necessary if flakes of sputtered material beginn to build and it has not to be a "perfect" cleaning process. Of course, you need to clean from time to time the viewing window, the guns, the penning gauge and the shutter.
If you have a vacuum problem, it is more likely that one of the o-rings in not OK. The most probable one is of course the rotating seal of the specimen mount. It should be cleaned and lubricated about every 50 hours of use.
Furthermore I can only encourage you to seek help from your local Gatan representative. At least the people from Gatan Germany never left us alone with our occasional problems and questions about the system and came up with help and solutions readily (hello Herr Nitschke, do you read this? :)
Petra
Usual disclaimer: I have no interest or relation to Gatan other than being a very satisfied customer.
At 12:21 01.10.99 +0900, you wrote: } } Dear members: } } I have used PIPS ion beam thinner for a month or so. Vacuum level became } bad and I could not turn on the gun. When I opened up the chamber, I found } that metal parts were contaminated severely. It seemed that it was time to } clean up the parts. } I tried to clean them with tissue paper and alcohol. They were hardly } removed. Especially when the metal surface had a machining marks such as } concentric circular grooves, it was more difficult. Some parts I could take } out of chamber and clean. The other parts, I could not take out because they } were stationary. I have to clean them in a state of being equipped. I am } afraid to use organic solvent which would remain in the chamber and make the } vacuum worse. } My question is how I can clean the contamination easily. Could any of you } give me help out of your good experiences? Help from Gatan expert would also } be appreciated. } } Jondo Yun } Center for Instrumental Analysis } Kyungnam University } 449 Weolyeong-dong, Masan, 631-701, Korea } 82-551-249-2697 (tel) } 82-551-248-5033 (fax) } jdyun-at-hanma.kyungnam.ac.kr
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
This position is for a technician in the Intel Fab 12 Materials Lab TEM group who will be responsible for TEM sample preparation. This job requires working with engineers to develop a sample preparation strategy and taking that task to completion. TEM sample preparation requires a high degree of manual skill, judgment, dexterity, hand-eye coordination, steady hands and good eyesight. The position requires operating precision mechanical sectioning and polishing equipment (tripod polisher and dimpler), focused ion beam (FIB) tools, and Ar-ion milling tools. High power optical microscopes and dual-beam FIBs are used to determine precision cross-section locations. TEM samples are extremely fragile and require extreme care and good manual dexterity during handling. Auxiliary duties will also include general upkeep of the TEM group labs, developing and printing TEM film and labeling TEM prints. The ideal candidate will have a 2-year college or technician degree in a science or engineering field and no less than 2-3 years experience operating TEM sample preparation equipment. The candidate will have worked in an area where manual dexterity, independent operation and decision making have been demonstrated. Familiarity with semiconductor process technology is also highly desirable. Intel is an equal opportunity employer. Resumes accepted by fax, email, or snail mail. Interested candidates should contact:
David Howell FAB 12 TEM Engineer Intel Corporation M/S OC2-218 4500 S. Dobson Rd Chandler, AZ 85248-4906 dave.fab12.howell-at-intel.com Fax (480)-715-8363
I now have a backup copy of the lost 3 days of messages courtesy of Kristian Ukkonen {kukkonen-at-cc.hut.fi} . I will simply append them to the archives for September when I update the Listserver archive site at http://www.msa.microscopy.com/MicroscopyListserver/MicroscopyArchives.html.
Thanks to everyone who offered to send me their copies of the Email for those 3 days . I'm please to see that there are other pack rats that also keep files for extended periods of time.
In a message dated 10/2/99 6:46:15 AM Eastern Daylight Time, zaluzec-at-Sparc5.Microscopy.Com writes:
{ { Subj: Administrivia: Nestor has backup copy of lost messages Date: 10/2/99 6:46:15 AM Eastern Daylight Time From: zaluzec-at-Sparc5.Microscopy.Com (Nestor J. Zaluzec) 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 On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Colleagues...
I now have a backup copy of the lost 3 days of messages courtesy of Kristian Ukkonen {kukkonen-at-cc.hut.fi} . I will simply append them to the archives for September when I update the Listserver archive site at http://www.msa.microscopy.com/MicroscopyListserver/MicroscopyArchives.html.
Thanks to everyone who offered to send me their copies of the Email for those 3 days . I'm please to see that there are other pack rats that also keep files for extended periods of time.
I agree with most of what you have said about the early mechanical advantages of Siemens and Philips over RCA, but I don't (and didn't) arrive at the same conclusion that using RCA was a big mistake. I did electron microscopy for my biological PhD thesis at Harvard about 1956-58 on an RCA EMU-2D. During my postdoc with Don Fawcett (at Cornell Med and then Harvard Med) in 1958-61 we used mostly RCA EMU-3. A Siemens IA was available to us during part of that time, but wasn't used much. As you pointed out, the long focal length of the RCA instruments was great for contrast, but less optimal for resolution. But why worry about only getting 7A resolution (instead of 4-5A) when the smallest structure of appreciable biological interest visible in EM sections was about 25A (sublayers of biological membranes). Was there any advantage in seeing the stain particles more sharply? On the other hand, contrast was extremely important, and contributed to some of the high quality EM produced by Fawcett and his associates.
When I left for my first faculty position (Anatomy Dept at Stanford Med, 1961-71), I had grant money for an EM, and picked an RCA EMU-3F. I never regretted that choice. The high contrast of that instrument was invaluable, especially when (beginning in 1966) I was developing a method and device for cutting ultrathin frozen sections (which were inherently low in contrast).
Although my later instruments were a Philips 300, JEOL 100B, and then Philips 201, Philips 400 and Philips CM10, I still look back on the RCA as a great instrument, despite its various inconveniences.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Anatomy and Cell Biology University of Michigan Medical School Ann Arbor, MI 48109-0616 Tel (work) 763-1287, Fax (work) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} Hello List Readers: } Laura Rhoads suggested that forced use of THAT instrument would be a } suitable penalty for those committing "crimes" such as broadcasting } "dogma". I concured, adding "I would not want to offend anybody, but } I've been in purgatory for two years already. Circa 1970 that instrument } was the worst money could buy in the Western World." } } Several readers have responded making positive points about that } instruments. I agree with some of these because nothing in this world is } entirely bad - even a broken clock shows the right time, twice a day. } } Yes, that instrument looked good, it had a big column and would make a } fine statue in someone's garden (so Laura suggested, personal } communications). I would love one, it would be the crowing glory to the } banks of the Ross River. } } Yes, the image had good contrast because it was a medium resolution } instrument (I guess about 0.8nm). A longer working distance objective } results in higher contrast but lower resolution. Other manufacturers } offer high contrast objectives, but most instrument purchasers ask for } higher resolution. } } Yes, RCA's are historical instrument and the first commercially produced } American TEM. Though I believe that Siemens produced the first commercial } TEM in the mid 30th. } } Yes, for some people this was their first electron microscope and it had } to be impressive to us who grew up before computer whizz-bangery. } Unfortunately, it was my fourth TEM. My previous loves had been a } mid-60th Siemens Elmiskop, a Philips 100C (the one with a near } horizontal column and transmission viewing screen) and a Zeiss 9C. All } of these were rather better suited to productive work. } } Yes, the electronics were reliable, but the vacuum gauging was } insufficient. Since it also had no vacuum locks and blanking provisions } were poor, vacuum trouble-shooting was very difficult. } } Yes, it was reasonably easy to operate and was great for forced coffee } breaks. No specimen, no gun, nor camera vacuum locks made for lots and } lots of pumping times. } } Yes, the camera had two options, either three single plates could be } inserted or one long plate taking five images on one plate. Bit of a } pain to share negatives with another operator. Absolutely awful for } taking lots of images and changing specimens. The pumping cycle was slow. } } The complete lack of vacuum locks and the poor film options made the } EMU-4 a "hysterical instrument". Such features were not rocket science } even in those days. More features . . . } } Changing the filament was a painful operation. I think the basic } alignment after filament change required three complete column pumping } cycles, lots of time for getting other jobs done during those } operations. I could not align that scope in under two hours. I had } inherited a service contract and the service men could do no better. } } Alignment screws were difficult to adjust accurately in those days on } most TEMs. I learned from the service man that the final touch was best } achieved by tapping the column with a rubber mallet - which was not part } of the service kit. } } Filament life averaged about 12 hours, with the alignment procedure } eating up a good part thereof. Short filament life may not have been an } inherent problem in the design. This may have been due to poor vacuum, } but there was no good way of isolating vacuum parts, getting a reliable } vacuum reading and finding a possible leak. } } } Why did RCA sell the EMU-4 to Forgflow, who then quickly discontinued } that instrument? } Because the column design was 10 years too late. By 1972 I had a Philips } EM300 - now that was progress. Philips at last had eclipsed the } Elmiskops, which had been the top brand throughout the 50th and 60th. } } Incidentally, Siemens closed their EM division about '78. In Karlsruhe in } October '77 I had admired Siemens' latest "Wunderkind", it turned out be } the Divisions death knell. It was a FESEM based somewhat on the research } by Crewe (Chicago ?) Nice instrument, beautiful column, but Vacuum } Generator of the UK had build a better performer at almost half the } price. . . . the rest is history. } } Disclaimer: Opinions expressed are mine they are based on fragile } memories. No dogma is implied or intended. Feel free to purchase an } EMU-4, if you can find one. Even Phantom comics of that period fetch } increasing prices and modern instruments depreciate rapidly. Don't } condemn me for another two years on THAT instrument please. PST does not } supply the EMU-4 or similar. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com
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How does the EMC-1 that I worked on fit into this product line? Do I have the model identification wrong? It *has* been over 30 years since I worked with the RCA 'scope.
gary g.
At 10:46 AM 10/2/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm pretty new at LM and I am trying to characterize components of seed cell walls. Does anyone know of stains that I could use to differentiate, say, celluloses from hemicelluloses from pectins etc.? I saw a really nice picture in a text once where different components were fluorescing at different colors, but I can't find the book now.
Also, if anyone knows of references to such experiments, I would be very grateful.
Thank you,
Dana Stewart Masters Student, Simon Fraser University British Columbia djstewar-at-sfu.ca
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Does anyone have a source for IR responsive marking pens? This is for use in a SEM with an IR chamber viewing camera system. The idea is to be able to mark specimen stubs and read numbers with the camera.
Low Vacuum and Environmental Scanning Electron Microscopy, LV-ESEM= '99
October 19-21, 1999, Chalmers University of Technology, Gothenburg,= Sweden
This course will be given in collaboration with four microscope manufacturers (Hitachi, Jeol, Leo, Philips/Fei) and suppliers of= equipment for EDX and EBSP.
The aim of this 3-day intensive course is to give a theoretical background in the morning sessions and experimental insights in the afternoons.= The lectures and the demonstrations will be given by application specialists from the different companies representad at the course and also by researchers working with LV- and ESEM. The demonstrations and lab= classes will be carried out on equipment brought to Chalmers specifically= for this course. =20
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Course organisers: Lena Falk (lklfalk-at-fy.chalmers.se) and Mats Halvarsson (mats.halvarsson-at-fy.chalmers.se)
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Associate Professor Lena Falk Department of Experimental Physics, Chalmers University of Technology, SE-412 96 G=F6teborg, SWEDEN
With regard to Jim Darley's citing of the High Crimes in the Diet of Dogma Court Record... } } Hello List Readers: } Laura Rhoads suggested that forced use of THAT instrument would be a suitable } penalty for those committing "crimes" such as broadcasting "dogma". I } concured, } adding "I would not want to offend anybody, but I've been in purgatory for } two } years already. Circa 1970 that instrument was the worst money could buy in } the } Western World." } } Yes, that instrument looked good, it had a big column and would make a fine } statue in someone's garden (so Laura suggested, personal communications). I } would love one, it would be the crowing glory to the banks of the Ross River.
Jim- You've got it all wrong: I said lawn ornament, not garden. If I put an EMU-4 column and requisite support pedestal in my garden there would be no room for my radishes... Actually, someone has already offered me an RCA EMT for this purpose, but that model isn't nearly massive enough for what I intend (I think it's complete so if anyone wants this artifact say, for a home workshop restoration project, contact me off-list).
Actually, the subject of RCA-for-pillory has brought up some rather interesting interpretations regarding the application of RCA EO equipment. Since I had yet to enter kindergarten by the time the last EMU-4 rolled off the assembly line in 1969 I never had the opportunity many others seem to have had cutting their teeth (I was cutting my own) on this (at the time) cutting edge technology. Missing this equipment in its prime, I feel somehow cheated, and as a result will have to rely on the experiences of those who did. What I find really interesting is the disparate variety of opinions as to service and operation. Could the EMU-4 Jim used at the World Expo have been a made on a Monday and Chuck Garber's, for example, a Wednesday? Or, did the RCA engineers install some extra parts they found in Hangar 18 from the Roswell, NM Crash Site that Jim's didn't get? Since the SPI rig is still running perhaps the operators can comment on their filament life and if it exceeds 12 hours? In order to solve the poor vacuum pumping problem (and to avoid forced coffee breaks) has this unit's vacuum system been retrofitted with a Balzers turbo pump perhaps? Are the alignment problems vividly recalled simply the result of using the wrong size mallet maybe?
All these issues are quite intriguing. If anyone does look into the history of RCA then maybe other manufacturers of EO equipment, such as Siemens, would be in order as well? And who was Vacuum Generator of the UK? Those who forget the past are doomed to repeat its failures. It seems that the technical information is available, and MSA has a tremendous body of institutional memory. It would be a shame to have this collective wisdom disappear.
For that matter, when I arrived at my latest job I discovered a complete Philips 75C hidden in the corner of the basement, with original warranty card (never filled out) which I am told still operates...Where will I ever get parts?...
************************************************ If the iron dice must roll, may God help us all...
Theobald von Bethmann-Hollweg German chancellor, August 1, 1914
************************************************ Laura Rhoads Biology Department SUNY Potsdam Potsdam, NY 13676 315-267-2260 315-267-3170 fax
I think this question is asked frequently but I haven't been able to locate any responses in the archives. I need to know what other labs charge for using the various pieces of equipment in your lab and for technical services performed by lab personnel. Thanks for your help.
Donna Wagahoff SIU School of Medicine PO Box 19627 Springfield, IL 62794-9627 217-782-0898 fax 217-524-3227
Kent: This thread was started as a jocular suggestion that the penalty for certain "sins" against the Listserver, should be forced labour on the RCA EMU-4. Similarly, I too used the subject header, "hysterical notes", also in the jocular. I have changed this to the proper term.
That previous contribution was in reply to three contributors and various points made. My own EM experience dates to the mid 60th and I had restricted my comments to instruments available from that time only.
I marveled then at the fine structure atlases produced by Fawcett and Rhodin and learned much from the technique book by Pease. I understand that prior to about 1962 there were at least three labs in USA, plus Sjostrand's in Sweden, another lab in the UK and the lab at Sydney University under Drummond, which were the only ones to consistently produce, even by today's standards, high quality images of tissues.
Good contrast was certainly due to longer objective lenses (and lower kV used), but in those days and that was before lead staining was introduced by Watson, section-contrast was actually better because the then used butyl-methyl methacrylates added much less electron density than do the epoxy resins. Those methacrylates had other problems, for sure, but contrast was a plus.
I doubt that all historically so successful labs used the particularly contrasty RCA TEMs, yet all were able to produce excellent images for various other reasons too.
I have never used the earlier models of RCA TEMs. I expect that they were good instruments in comparison with others on the market at the time.
Chuck Garber evidently has a "soft spot" for the EMU-4 and had wondered why that model was not successful in the marked place and had lead to RCA's withdrawal from the field. I think that the reason for poor sales are obvious: by the late 60th there were several very competent instruments available, some had high contrast objectives too, but the crucial advantages of those instru ments were vacuum locks for specimen, for filament and for film changes. Also, some manufacturers realized that memories were not enough, people had to be able to take lots of photographs. RCA had underestimated the importance of all these features and I believe these were the reasons for the demise of the RCA TEMs. "Yankee" ingenuity is famous and unquestioned, rather the EMU-4 is an example of a manufacturer not watching the opposition and a failure to listen to customers increased requirements. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Sunday, October 03, 1999 12:47 AM, A. K. Christensen [SMTP:akc-at-umich.edu] wrote:
} Jim, } } I agree with most of what you have said about the early mechanical } advantages of Siemens and Philips over RCA, but I don't (and didn't) arrive } at the same conclusion that using RCA was a big mistake. I did electron } microscopy for my biological PhD thesis at Harvard about 1956-58 on an RCA } EMU-2D. During my postdoc with Don Fawcett (at Cornell Med and then } Harvard Med) in 1958-61 we used mostly RCA EMU-3. A Siemens IA was } available to us during part of that time, but wasn't used much. As you } pointed out, the long focal length of the RCA instruments was great for } contrast, but less optimal for resolution. But why worry about only } getting 7A resolution (instead of 4-5A) when the smallest structure of } appreciable biological interest visible in EM sections was about 25A } (sublayers of biological membranes). Was there any advantage in seeing the } stain particles more sharply? On the other hand, contrast was extremely } important, and contributed to some of the high quality EM produced by } Fawcett and his associates. } } When I left for my first faculty position (Anatomy Dept at Stanford Med, } 1961-71), I had grant money for an EM, and picked an RCA EMU-3F. I never } regretted that choice. The high contrast of that instrument was } invaluable, especially when (beginning in 1966) I was developing a method } and device for cutting ultrathin frozen sections (which were inherently low } in contrast). } } Although my later instruments were a Philips 300, JEOL 100B, and then } Philips 201, Philips 400 and Philips CM10, I still look back on the RCA as } a great instrument, despite its various inconveniences. } } Kent } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } A. Kent Christensen, Professor Emeritus } Department of Anatomy and Cell Biology } University of Michigan Medical School } Ann Arbor, MI 48109-0616 } Tel (work) 763-1287, Fax (work) 763-1166 } akc-at-umich.edu } http://www.umich.edu/~akc/ } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } --On Mon, Sep 27, 1999 9:50 PM +1000 jim {jim-at-proscitech.com.au} wrote: } } } Hello List Readers: } } Laura Rhoads suggested that forced use of THAT instrument would be a } } suitable penalty for those committing "crimes" such as broadcasting } } "dogma". I concured, adding "I would not want to offend anybody, but } } I've been in purgatory for two years already. Circa 1970 that instrument } } was the worst money could buy in the Western World." } } } } Several readers have responded making positive points about that } } instruments. I agree with some of these because nothing in this world is } } entirely bad - even a broken clock shows the right time, twice a day. } } } } Yes, that instrument looked good, it had a big column and would make a } } fine statue in someone's garden (so Laura suggested, personal } } communications). I would love one, it would be the crowing glory to the } } banks of the Ross River. } } } } Yes, the image had good contrast because it was a medium resolution } } instrument (I guess about 0.8nm). A longer working distance objective } } results in higher contrast but lower resolution. Other manufacturers } } offer high contrast objectives, but most instrument purchasers ask for } } higher resolution. } } } } Yes, RCA's are historical instrument and the first commercially produced } } American TEM. Though I believe that Siemens produced the first commercial } } TEM in the mid 30th. } } } } Yes, for some people this was their first electron microscope and it had } } to be impressive to us who grew up before computer whizz-bangery. } } Unfortunately, it was my fourth TEM. My previous loves had been a } } mid-60th Siemens Elmiskop, a Philips 100C (the one with a near } } horizontal column and transmission viewing screen) and a Zeiss 9C. All } } of these were rather better suited to productive work. } } } } Yes, the electronics were reliable, but the vacuum gauging was } } insufficient. Since it also had no vacuum locks and blanking provisions } } were poor, vacuum trouble-shooting was very difficult. } } } } Yes, it was reasonably easy to operate and was great for forced coffee } } breaks. No specimen, no gun, nor camera vacuum locks made for lots and } } lots of pumping times. } } } } Yes, the camera had two options, either three single plates could be } } inserted or one long plate taking five images on one plate. Bit of a } } pain to share negatives with another operator. Absolutely awful for } } taking lots of images and changing specimens. The pumping cycle was slow. } } } } The complete lack of vacuum locks and the poor film options made the } } EMU-4 a "hysterical instrument". Such features were not rocket science } } even in those days. More features . . . } } } } Changing the filament was a painful operation. I think the basic } } alignment after filament change required three complete column pumping } } cycles, lots of time for getting other jobs done during those } } operations. I could not align that scope in under two hours. I had } } inherited a service contract and the service men could do no better. } } } } Alignment screws were difficult to adjust accurately in those days on } } most TEMs. I learned from the service man that the final touch was best } } achieved by tapping the column with a rubber mallet - which was not part } } of the service kit. } } } } Filament life averaged about 12 hours, with the alignment procedure } } eating up a good part thereof. Short filament life may not have been an } } inherent problem in the design. This may have been due to poor vacuum, } } but there was no good way of isolating vacuum parts, getting a reliable } } vacuum reading and finding a possible leak. } } } } } } Why did RCA sell the EMU-4 to Forgflow, who then quickly discontinued } } that instrument? } } Because the column design was 10 years too late. By 1972 I had a Philips } } EM300 - now that was progress. Philips at last had eclipsed the } } Elmiskops, which had been the top brand throughout the 50th and 60th. } } } } Incidentally, Siemens closed their EM division about '78. In Karlsruhe in } } October '77 I had admired Siemens' latest "Wunderkind", it turned out be } } the Divisions death knell. It was a FESEM based somewhat on the research } } by Crewe (Chicago ?) Nice instrument, beautiful column, but Vacuum } } Generator of the UK had build a better performer at almost half the } } price. . . . the rest is history. } } } } Disclaimer: Opinions expressed are mine they are based on fragile } } memories. No dogma is implied or intended. Feel free to purchase an } } EMU-4, if you can find one. Even Phantom comics of that period fetch } } increasing prices and modern instruments depreciate rapidly. Don't } } condemn me for another two years on THAT instrument please. PST does not } } supply the EMU-4 or similar. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } }
A 2-year postdoctoral research fellowship is available to characterise radiation damage induced by neutron irradiation in model alloys and ferritic steels through the use of advanced transmission electron microscopy. The project aim is to determine the nature and the compositional dependency of the damage and, hence, to contribute to the understanding of the decrease in fracture toughness with neutron exposure in RPV steels. The successful candidate will work closely with industrial collaborators during the course of the project. Please quote ref: JMT1 in any correspondence.
The project requires the systematic examination of specimens from a range of model alloys subjected to different thermal and irradiation treatments to characterise the matrix damage present in the form of point defect clusters and dislocation loops, using weak beam analysis in energy-filtered images. In addition, possible interactions between point defects and alloying elements will be explored using analytical methods including: core-loss imaging, electron energy-loss spectroscopy and EDX analysis. The Department of Materials operates a range of electron microscopes including a JEOL 3000F FEG-TEM with GIF and EDX, an HB501 dedicated FEG-STEM and 400keV HREM instruments. The successful applicant will join a rapidly growing group of post-doctoral and postgraduate students engaged in microstructural and chemical characterisation of a wide range of materials using electron microscopy.
The successful candidate will possess a PhD in a materials science, physics or materials engineering field, be able to work within a group and be able to liase with industrial collaborators. Experience using transmission electron microscopy for the characterisation of crystal defects is essential.
2. Postdoctoral Fellowship in SCC/Electron Microscopy
A 2-year postdoctoral research fellowship is available to develop the use of analytical transmission electron microscopy methods to investigate environmentally assisted cracking (EAC) in austenitic stainless steels, in collaboration with Rolls-Royce Marine Power. Please quote ref: JMT2 in any correspondence.
The post concerns: (i) the development of specimen preparation techniques for generation of electron-transparent foils from materials containing cracks; and (ii) the systematic examination of a series of materials of selected compositions in which EAC has been induced in aqueous environments under controlled stress, temperature, environmental chemistry and corrosion potential. Characterisation will be performed using a range of electron optical equipment including a new JEOL 300kV FEG-TEM and an HB501 FEG-STEM. The applicant will join a rapidly growing group of post-doctoral and postgraduate students engaged in structural and chemical characterisation of a wide range of materials using electron beam methods.
Candidates should possess a PhD in a materials science, physics or materials engineering field, be able to work within a group and be able to liase with industrial collaborators.
We obtain our EM images using 35mm film. Each roll of film normally has about 60 images. We would like to eliminate the cost of printing these images and work with digital images. We have considered cutting our rolls of negatives into short strips and scanning them on a flatbed scanner. We have rejected this idea because of the time needed to digitally cut and save each individual image from the huge original image. We have also considered cutting our rolls into 6 image strips and scanning them with a film scanner. We have rejected this idea because the only film scanners we have found can only read strips with a maximum of 6 images making it necessary to load a new 6 image strip every few minutes. Again not a very efficient method.
Does anyone know of a film scanner that will read 35mm strips of 20 or more images or have some other idea how to automate the scanning of our negatives?
John
John M. Basgen Department of Pediatrics University of Minnesota Box 491 420 Delaware Street SE Minneapolis, MN 55455 Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-tc.umn.edu
The Center for High Resolution Electron Microscopy at Arizona State University has an open position for a Research Specialist. The Center provides electron microscopy resources for the university and external community, including a range of transmission and scanning electron microscopes, as well as specimen preparation facilities. The successful applicant will be expected to assist with the day-to-day operation and maintenance of microscopes within the Center. Primary instrument responsibility will include but not be limited to the JEOL 2000FX transmission electron microscope (TEM), the JEOL 840 Scanning EM, and the soon-to-be acquired JEOL 5900 SEM with liquid-helium CL system. Other tasks will include training and supervision of researchers in proper use of the instruments, including observance of safety procedures, and authorization of users. Some limited opportunity to assist and advise researchers in designing and carrying out experiments, including active participation and collaboration in experiments with data collection and analysis as required.
Minimum qualifications: Bachelors or Masters degree or equivalent training in Physical or Materials' Sciences or closely related discipline, with at least three but preferably five years' additional experience with operation and maintenance of scanning and/or transmission electron microscopes. Experience with design, construction and testing of electronic circuitry, and knowledge of vacuum systems would be desirable.
Application deadline: October 15, 1999, or until position filled.
Arizona State University is an equal opportunity employer. Women and minorities are encouraged to apply.
Send resume and the names and addresses of three references to John Wheatley. Please contact John via one of the following ways--preferably e-mail:
John C. Wheatley, Lab Manager Arizona State University Center for Solid State Science Tempe, AZ 85287-1704
In my earlier message I didn't make myself clear about what I actually need. The bean counters here are wanting to raise the rates that we charge in-house people to use the equipment and to provide technical help to those who need it. I need to know what other state facilities charge for these services to justify our raising our rates or keeping them where they are. Thanks for your help.
Donna Wagahoff SIU School of Medicine PO Box 19627 Springfield, IL 62794-9627 217-782-0898 fax 217-524-3227
To get an idea about the fee structure (to be revised soon) CIMC uses for services within our university, please visit our website and go to the fees page. (http://www.cimc.cornell.edu/Pages/fee.htm)
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} } } } I think this question is asked frequently but I haven't been able to } locate any responses in the archives. I need to know what other labs } charge for using the various pieces of equipment in your lab and for } technical services performed by lab personnel. Thanks for your help. } } Donna Wagahoff } SIU School of Medicine } PO Box 19627 } Springfield, IL 62794-9627 } 217-782-0898 } fax 217-524-3227
Hope this is the start of a great new week for everyone! My question today= is again delicate, but in a different way. I'm not even sure that it can be answered directly given a previous thread on commercialism, price fixing, undercutting the private sector with taxpayer funded instrumentation, unfair competetion on other levels, etc., but I'm going to ask anyway and bravely accept any criticism or flames if I don't phrase it correctly. I hope I'm= not exasperating anyone.=20
We have a Balzer's HPM010 high pressure freezer in our facility, one of= about 10 in the US if I'm not wrong. I actually know that there is a user's group out there, but a HD crash a bit back removed all my files on that, so I'll use= the list at least once to ask my question. I realize that MOST other labs must charge all users for instrument use based on beam time, number of cycles, or some such. We may be unique in that we do not charge anyone except off= campus or out of system users for our equipment. Each user supplies all= comsumables, the facility Supervisor and manager (me) are line items in our department's budget, and our service contracts are supported by our serviced departments, the College, and the Vice-provost for research. We consider ourselves extremely lucky not to have to charge for use of the lab so that even unfunded people can work here if they can get supplies and we don't have to do the billing and accounting associated with charges. We diligently try to maintain excellent relations with the commercial EM labs around our area and refer all business to them if they have the facilities to perform it. Hence, we don't have much experience in setting prices for users that we can charge. How do the rest= of you decide on these charges? I have met with our comptroller's office (a couple of years ago when we were looking at charging fees like all the rest of the labs on campus) and understand that if you charge anyone with a government contract or grant, you must bill EVERYONE, even for teaching time, and that the cost to grantees or contract=A0 holders must be billed at the lowest rate.= Costs must be neutral - that is, no profit and all actual costs recovered, which means a lot of research into just what those costs ARE. It is very difficult for an essentially one man band to do all this stuff and it was decided to leave us alone as we are for now.=20
Hence, my problem. (FINALLY).=A0 Can I get some idea of what others charge= for High Pressure Freezing=A0 without making every service provider out there= angry and being accused of unfair competition? Or do I just guess for a one time industrial user who may not really want what we can do, anyway? I have, in= the past, charged $5 per "shot" plus a Dewar of liquid nitrogen at cost to cover expendibles and time, this to outside but not for profit users - these were visiting scientists from around the world and here in state. I'm not sure= this actually covered our costs.=20
Any help or guidance would be useful and appreciated. Any criticism of my= post (to include spelling or attitude) will be greatfully received and= considered. Thanks in advance.
William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899 =20
I am faced with my frequent dilemma of trying to look at a sample= I have never seen before, and one that my customer has either never imaged before or has not provided images of the desired features. Has anyone ou= t there ever imaged the materials used in soft contact lens in the SEM before? If so, are there any things that might help to either enhance or preserve the fine structure of the material they are made from? I am happ= y for any suggestions or advice. Thanks in advance for any input.
Dear Listers, We are interested in doing the freeze fracture/freeze etch procedure I did years ago on a Balzers instrument. I have two questions: a) are any of you using this technique and if so what model are you using? b) will you accept contract work? Thanks in advance for your assistance. Rosemary
I posted this last week, but have had only a couple of replies. Could it, and its successor of Thurs 30th have gotten caught up in the problems? Plse advise so that I can re-post if neccessary.
I had a nice time at UC Davis last month, packing up the 840 we bought from them, now all I have to do is to galvanise our admin into commissioning some miniscule room alterations (you can read that both ways), and she'll be a cracker, cobber!
Dear all, Iwould first like to thank everyone who provide information on how to measure fingerprints. On a different matter I intend to fire up a UV laser (emmitting at 248nm) for the first time in several years in the near future. I am obviously uncertain that the laser will work and do not wish to spend too much time trying to optimise the laser and so on without being absolutely sure it is, in fact, lasing. For this reason I was wandering if anyone could recomend a fluorescent dye (or other material) that I could place in the beam path which would emit in the visible region and thus enable me to be sure of the laser's efficacy. Thanks in advance. -------------------------------------------------------------------------------- -----------------------Dr. Giles SandersZeneca / SmithKline Beecham Centre for Analytical SciencesChemistry DepartmentImperial College of Science, Technology and MedicineLondonSW7 2AY----------------------------------------------------------------------------- -----------------------
I'm in the process of fitting the factory optical microscope onto an 840A, and I don't have any info on the various cable connections.
1 There are two cables issuing from the mounting flange:
- one with a BNC plug, cable labelled "OM1" - one with a 5-pin plug, cable labelled "BE8"
2 There's one cable issuing from the elbow, with a 5-pin plug, and a smudged label.
3 On the elbow, there's a 5-pin socket, also what looks like a little green indicator light.
I will be very grateful to whoever can explain what all of these are for (they are in addition to the 5-pin plug from the illumination lamp base).
Also, does the secondary electron imaging still work with the OM fitted? Presumably the latter has to be thoroughly retracted.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
We are setting up a microscopy techniques require an environmental chamber with carbon dioxide control within 5% +/- 0.5%. Our microscope system is Zeiss Axiovert 135. Does anybody know the supplier, or have experience to build one up? We need your help.
Some fifteen years ago I ran a course for a contact lens manufacturer where we modified their microscope (SEM) to look at the wet lens.
They had an old ISI SEM but complete with a Robinson BSE detector. As the SEM had a "proper" vacuum system, with a manifold leading from the DP to the specimen chamber and gun area, we were able to modify the system to be pseudo VP; I have to say in those days we just called it common sense!
We placed a rubber bung (stopper to some) in the rear pumping line of the specimen area. The bung had a 0.5cm (3/16th inch) hole in it. Switching OFF all voltages to the SE (Everhart Thornley) detector we than ran in backscatter for about 20 minutes each session, by which time the lens would start to curl. We did not fix the lens down simply placed it on a 1 1/.4 inch stub.
The porosity in the lens was very clear under these conditions.
Hope this helps?
Steve Chapman
Senior Consultant E.M. Protrain, 6 Hillcrest Way, Buckingham Industrial Park, Buckingham MK18 1FU, England Tel & Fax 44 (0)1280 814774 E-mail - protrain-at-emcourses.com Web Site - http://www.emcourses.com For Consultancy and Courses in Electron Microscopy World Wide Courses available in - Australia, Canada, Europe, South Africa, New Zealand, Taiwan, United States, United Kingdom
We are setting up a microscopy techniques require an environmental chamber with carbon dioxide control within 5% +/- 0.5%. Our microscope system is Zeiss Axiovert 135. Does anybody know the supplier, or have experience to build one up? We need your help.
Greetings, I am in the market to buy a stereo microscope with a price of up to about $3000. One major use of the microscope will be to examine snail shells for defects at high magnifications, so as good a resolution as $3000 can buy is the major criterion. The two lines I am considering are the Olympus SZ & the Leica GZ series. I don't have the means of personally testing different models. I will appreciate comments from those who have used these models on the optics, mechanics & any other aspect of these microscopes. Thanks.
Greetings, I am in the market to buy a stereo microscope with a price of up to about $3000. One major use of the microscope will be to examine snail shells for defects at high magnifications, so as good a resolution as $3000 can buy is the major criterion. The two lines I am considering are the Olympus SZ & the Leica GZ series. I don't have the means of personally testing different models. I will appreciate comments from those who have used these models on the optics, mechanics & any other aspect of these microscopes. Thanks.
I have some negatives taken on the TEM which developed extremely light. I was wondering if anyone knows any tricks on photoshop for generating good quality prints. I typically adjust the levels and condense the size of the picture, but I am still not completely happy about the quality of the prints.
Looking at the post of Michael D. Frey, about sample preparation for contact lens, I reflexioned that this is a very good place to ask you all ideas for service with SEM, This is the unique SEM in our State, even I know that only there is another one in Southeast Region of Mexico, including our neighboard country of Guatemala. So I would like to extend my work in investigation assistance in Biology Science, to another services to customers for different aplications..... I«ve known some forensic applications, but I also know that this services needs besides me, to operate the SEM, one expert who knows what to look at the sample..
Your opinion will be very helpfull to solve this dilema of how to offer services to customers
We would like to buy immediately used GIFs for 300kV and 120kV TEMs.
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories (MIL) and Department of Biology University of California at San Diego
address: 1500 Bonner Hall, University of California at San Diego 9500 Gilman Drive, La Jolla, CA 92093-0368 telephone - lab: 8585342484 telephone - office: 8588223373 pager: 8586161420 fax: 8588223715 email: mmm-at-ucsd.edu www site: http://mil.ucsd.edu
Need some extra stages? Always wanted to do complimentary replicas? Now's your chance. I've gotten out of the freeze fracture business and put my old Balzers Turbo out to pasture. I did, however, save a hoard of parts which someone might put to good use. 10 quartz crystals 3 3 position specimen tables for counterflow loading system, plus loading rods 80 3mm gold specimen carriers for above 2 4 position specimen tables 1 1 position specimen table 1 complimentary (double) replica table plus lots of gold and Cu plates 1 specimen table for monolayer tissue 10 tubes of tungsten filaments for e guns 16 tubes carbon rods fo e gun 20 pre-drilled C rods for Pt bullets 55 Pt bullets 1/2 box Balzers microtome blades 2 12 well porcelain spot plates T/W wire and rods for evaporation Gordon Stereoscope (front silvered mirrors, adjustable 10x eyepieces) for examining stereo EM (up to 8X10 side by side) TONS of em grids of every possible mesh and configuration
Contact me if you're interested. This offering goes as a package, not individual items.
We would like to buy immediately used GIFs for 300kV and 120kV TEMs.
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories (MIL) and Department of Biology University of California at San Diego
address: 1500 Bonner Hall, University of California at San Diego 9500 Gilman Drive, La Jolla, CA 92093-0368 telephone - lab: 8585342484 telephone - office: 8588223373 pager: 8586161420 fax: 8588223715 email: mmm-at-ucsd.edu www site: http://mil.ucsd.edu
Dear David, I have looked at a soft contact lens before, but just in the dried state. I just stuck it on a stub and gold coated it. Since they are normally 55% water, I wouldn't vouch for the stucture in this state. Being smooth jelly, I don't think they have much surface structure. At 03:23 PM 10/4/99 -0400, you wrote:
} } Dear Listserver Members, } } I am faced with my frequent dilemma of trying to look at a sample I } have never seen before, and one that my customer has either never imaged } before or has not provided images of the desired features. Has anyone out } there ever imaged the materials used in soft contact lens in the SEM } before? If so, are there any things that might help to either enhance or } preserve the fine structure of the material they are made from? I am happy } for any suggestions or advice. Thanks in advance for any input. } } David Frey } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
At 11:20 AM 10/5/99 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Rosemary,
We have a BALZERS BAF060 freeze fracture machine. This is the newest model of this kind of instrument. Please chcke our web site (http://www.cbc.umn.edu/em/) for more information.
Ya Chen Scientist
EM Facility Dept. of Genetics, Cell Biology, and Development University of Minnesota Medical School 6-160 Jacson Hall 321Church St. S.E. Minneapolis, MN 55455
I am looking for some information on using SEM in archaeology, ie, methods which might be used to determine what clay archaeological pieces were used for. Is there an on-line search site (like Medline in the Biological Sciences) in which I can type in keywords and come up with Journal articles? Anyone ever do something like this? Any information would be greatly appreciated.
I am looking for some information on using SEM in archaeology, ie, methods which might be used to determine what clay archaeological pieces were used for. Is there an on-line search site (like Medline in the Biological Sciences) in which I can type in keywords and come up with Journal articles? Anyone ever do something like this? Any information would be greatly appreciated.
Photoshop processing can help, but the place to start is in scanning. If you have a 30-bit or better yet a 36-bit (color) scanner, you should be able to manipulate the scanner's tone controls (black, white, and gamma) to optimize the scanner's 8-bit (grayscale) output. You might also try superimposing a neutral density filter on the film negative to match the work density to the scanner's range. The filter can be another piece of light-exposed and developed TEM film. It just needs to have a uniform (gray) background. Always scan TEM negatives as positive transparencies; then invert the contrast in the scanner or Photoshop.
A work-around, especially if you don't have a good scanner, is to print the negative in the darkroom to get the contrast and density you want and then scan the print.
To further adjust the tonal quality in Photoshop, use Levels or Curves adjustment layers rather than applying these commands directly. This will avoid much of the data loss in applying multiple corrections. Use the Multiply blending mode to increase the contrast from a light negative, and apply as many layers as needed to get the contrast and background brightness right for your printer. If you use unsharp mask filtering (highly recommended after scanning) and background leveling, apply these to layers before the tonal adjustments. Learning to use Layers in Photoshop takes some time, but is highly worthwhile. Good luck.
Larry Thomas
Larry Thomas Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto: Larry.Thomas-at-pnl.gov
From: Donald Delaney Sent: Tuesday, October 5, 1999 6:49 AM To: microscopy-at-Sparc5.Microscopy.Com Subject: Photoshop Negative Processing
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
I have some negatives taken on the TEM which developed extremely light. I was wondering if anyone knows any tricks on photoshop for generating good quality prints. I typically adjust the levels and condense the size of the picture, but I am still not completely happy about the quality of the prints.
We have a Kevex Quantum UTW that we have been using for over 10 years. We had to send it in once for window repair after one of the miniscule panes in the window sprang a leak. It has been fine since.
We also have a Oxford Link ISIS Ge detector with UTW. Seems like it has been around for 5 years with no problem.
Basically, we have found them to be no particular bother and well worth having in order to see the C and O peaks.
Warren
At 08:11 PM 10/4/1999 -0500, you wrote: } From: Self {GLGNOV2/RSIMS} } To: Microscopy-at-sparc5.microscopy.com } Subject: Ultrathin Window vs Be } Date: Wed, 29 Sep 1999 14:48:43 GMT+1200 } } Hi, Everyone } } I'm shortly going to buy a new EDS detector. } I'd quite like the performance advantages of a 1-atmosphere UTW, but } I'm concerned about the durability and longevity compared with } standard Be windows. } } Anyone out there got any relevant experience/views? } } Either to the list, or directly to me, please. } } thanks } } Ritchie
I am looking for some information on using SEM in archaeology, ie, methods which might be used to determine what clay archaeological pieces were used for. Is there an on-line search site (like Medline in the Biological Sciences) in which I can type in keywords and come up with Journal articles? Anyone ever do something like this? Any information would be greatly appreciated.
I have 20 new filaments; several Wehnelt assemblies; SEI, TE, CL?, detectors; Scintillator disks; and other small items from an ISI Model 40 SEM. They are free if someone can use them. Please get in touch with me directly via email at probe-at-mailbox.cc.binghamton.edu or phone 607 777-2832.
Best Regards, Bill Blackburn Geology Department Binghamton University Binghamton, NY 13902-6000
Dear Listers: I am running an SEM in the Geoscience Department at UNLV. I have been approached by a member of the biology department, who would like to image bacteria, the CaCO3 crystals deposited by the bacteria and if possible the substrate. The bacteria are fixed in glutaraldehyde. Does someone out there have a simple protocol for preparation and coating of these types of samples. I would like to apologize in advance for asking such a simple question, but I am only a geologist... :)
Sarah A. W. Lundberg Microbeam Facility Analyst Office (702) 895-1134 University of Nevada, Las Vegas Lab (702) 895-2660 4505 Maryland Parkway Box 454010 email lundberg-at-nevada.edu Las Vegas, NV 89154-4010 Fax (702) 895-4064
Home : 4489 De Forest Street Las Vegas, NV 89103 (702) 871-9635
Folks, I would like to pass on a message from a colleage which may be of interest to users of a SEM ISI model 40:
-----Original Message--------------------- } From: probe-at-mailbox.cc.binghamton.edu [mailto:probe-at-mailbox.cc.binghamton.edu] Sent: Tuesday, October 05, 1999 2:26 PM To: heichelb-at-binghamton.edu
Folks, I would like to pass on a message from a colleage which may be of interest to users of a SEM ISI model 40:
-----Original Message--------------------- } From: probe-at-mailbox.cc.binghamton.edu [mailto:probe-at-mailbox.cc.binghamton.edu] Sent: Tuesday, October 05, 1999 2:26 PM To: heichelb-at-binghamton.edu
Hello All,
We have a problem with the cooling water on a JEOL 1200EX TEM, and hope that someone can suggest a cause.
The instrument was run for years on a central closed-circuit water chiller which was also feeding several other instruments. About three years ago the chiller was replaced with a new central unit, and it was then the problem started. Over a period of about three weeks the water turns a dark brown to such an extent that it impossible to see through the sight glasses on the flow meters. The brown deposit has been analysed on an EDAX system, and has been shown to be rust. This is further borne out by the fact that the deposit can easily be removed from the flow meters using phosphoric acid.
Thinking that the problem was tied up with the new central chiller, we have now purchased a new chiller unit solely to cool the JEOL. We are using tap water in the system as recommended by the chiller manufacturer, and no additives are being used. The water circuit external to the TEM contains only copper, brass and plastic, and yet the brown rust deposit is still forming just as rapidly as before. Obviously we are very worried that there is some iron component in the TEM itself which is corroding away, and we could be faced with an expensive repair. We would be very interested to hear if anyone else has experienced this problem with a 1200EX.
Regards to all. Bob Phillips ****************** MicroServiS, Huntingdon, Cambs. UK *******************
I do not have on my hands the same microscope with spectrometers, therefore I will attempt to answer using circuit diagrams. 1. Backscattered electrons move with a high speed and along linear trajectories, therefore any hindrance on their ways produces the shadowing of BSE detector, which is on polepiece of objective lens, and BSE picture loss. The inserted optical microscope is such hindrance in your case. To take the BSE image with inserted optical microscope, there is an additional BSE detector on lower side of the optical microscope, which should connected with BSE preamplifier through the plug BE8 instead of the former detector . You should have also blank socket BE8 for connecting of unused detector to ground. During operation with spectrometers the high voltage feeds electrostatic electron trap placed on optical microscope through the plug OM1. This plug should be connected through a cable with the socket HV1 of SMXA - HVPS40 unit. 2. I think, this cable is labelled OMC4, it should be connected to the appropriate plug on OM CONTROL UNIT. 3. Illumination lamp of the microscope is feeded by this socket, therefore the cable OM on the base of illumination lamp should be inserted into this socket. The detector of secondary electrons will work with inserted optical microscope, but the picture will be worse even because for inserting of the optical microscope you should increase a working distance. But to avoid PMT destroying you can not turn on the illumination lamp of the microscope simultaneously with SEI detector. Hope this will help.
Best regards.
Victor Sidorenko, ANTRON Co. Ltd., scientific service, Moscow, Russia.
} Hi } } I'm in the process of fitting the factory optical microscope onto an 840A, } and I don't have any info on the various cable connections. } } 1 There are two cables issuing from the mounting flange: } } - one with a BNC plug, cable labelled "OM1" } - one with a 5-pin plug, cable labelled "BE8" } } 2 There's one cable issuing from the elbow, with a 5-pin plug, and a } smudged label. } } 3 On the elbow, there's a 5-pin socket, also what looks like a } little green indicator light. } } I will be very grateful to whoever can explain what all of these are } for (they are in addition to the 5-pin plug from the illumination } lamp base). } } Also, does the secondary electron imaging still work with the OM } fitted? Presumably the latter has to be thoroughly retracted. } } thanks } } Ritchie } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
Steve - I made a very similar modification to an Etec Autoscan at about that time. It was not a simple modification because we needed it to be quickly reversible and the Etec vacuum manifold complicated the low vacuum modification quite a lot. It worked very well for material specimens including atomic number contrast of uncoated mineral sections, uncut rock samples and any other samples that had low water contents, including insects.
Many of the readers are biologists and they should be aware that such a modification, which amounts to mechanical pump vacuum in the specimen chamber with near normal diffusion pump pressure in the gun chamber. It is only useful to about 2000x and useless for really wet tissue e.g. a piece of liver. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, October 05, 1999 5:55 PM, Steve Chapman [SMTP:PROTRAIN-at-CompuServe.COM] wrote:
} Hi,
} Some fifteen years ago I ran a course for a contact lens manufacturer where } we modified their microscope (SEM) to look at the wet lens. } } They had an old ISI SEM but complete with a Robinson BSE detector. As the } SEM had a "proper" vacuum system, with a manifold leading from the DP to } the specimen chamber and gun area, we were able to modify the system to be } pseudo VP; I have to say in those days we just called it common sense! } } We placed a rubber bung (stopper to some) in the rear pumping line of the } specimen area. The bung had a 0.5cm (3/16th inch) hole in it. Switching } OFF all voltages to the SE (Everhart Thornley) detector we than ran in } backscatter for about 20 minutes each session, by which time the lens would } start to curl. We did not fix the lens down simply placed it on a 1 1/.4 } inch stub. } } The porosity in the lens was very clear under these conditions. } } Hope this helps? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 6 Hillcrest Way, Buckingham Industrial Park, Buckingham MK18 1FU, } England } Tel & Fax 44 (0)1280 814774 } E-mail - protrain-at-emcourses.com } Web Site - http://www.emcourses.com } For Consultancy and Courses in Electron Microscopy World Wide } Courses available in - Australia, Canada, Europe, South Africa, New } Zealand, Taiwan, United States, United Kingdom
It's obviously a little late to be asking about this, since the baby is fine and almost 10 months old... BUT I was wondering if anyone has any info on using an SEM while pregnant? I used the one here at SUNY Potsdam during my first and third trimesters, as well as during the baby's first 5 months. I keep having these awful scenarios running through my mind about the old "Incredible Hulk" show where the guy was exposed to gamma rays and became the Hulk!
The main problem is that if the information is not in the neg, it cannot be created using Photoshop. It can be subtracted or contrast enhanced but not created. It is always better to overexpose whenever deciding which way to go.
Also, scanners tend to produce better results when set to negative rather than positive. This is because the neg has an inherently greater tonal range than a positive and the scanner tries to capture that. If you have a neg, scan as a neg. I have always achieved better results this way.
Printing an underexposed neg is going to produce a contrasty print. Still no additional info. Actually less. The only viable method of fixing an underexposed neg is to re-shoot it.
gary g.
At 03:15 PM 10/5/99 , you wrote:
} Photoshop processing can help, but the place to start is in scanning. If you } have a 30-bit or better yet a 36-bit (color) scanner, you should be able to } manipulate the scanner's tone controls (black, white, and gamma) to optimize the } scanner's 8-bit (grayscale) output. You might also try superimposing a neutral } density filter on the film negative to match the work density to the scanner's } range. The filter can be another piece of light-exposed and developed TEM } film. It just needs to have a uniform (gray) background. Always scan TEM } negatives as positive transparencies; then invert the contrast in the scanner or } Photoshop. } } A work-around, especially if you don't have a good scanner, is to print the } negative in the darkroom to get the contrast and density you want and then scan } the print. } } To further adjust the tonal quality in Photoshop, use Levels or Curves } adjustment layers rather than applying these commands directly. This will avoid } much of the data loss in applying multiple corrections. Use the Multiply } blending mode to increase the contrast from a light negative, and apply as many } layers as needed to get the contrast and background brightness right for your } printer. If you use unsharp mask filtering (highly recommended after scanning) } and background leveling, apply these to layers before the tonal adjustments. } Learning to use Layers in Photoshop takes some time, but is highly worthwhile. } Good luck. } } Larry Thomas } } Larry Thomas } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto: Larry.Thomas-at-pnl.gov } } } } } From: Donald Delaney } Sent: Tuesday, October 5, 1999 6:49 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Photoshop Negative Processing } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have some negatives taken on the TEM which developed extremely light. } I } was wondering if anyone knows any tricks on photoshop for generating } good } quality prints. I typically adjust the levels and condense the size of } the } picture, but I am still not completely happy about the quality of the } prints. } } Don Delaney } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Yes, It's true the gamma radiation mutates the DNA after it passes threough 2 inches of steel.
The symtoms only become apparent after the child is in his teenage years and subsides after age 21.
Ask any parent of an SEM operator who has teenage kids.
Earl
Randy & Jenna & Orin Brown wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } It's obviously a little late to be asking about this, since the baby } is fine and almost 10 months old... BUT I was wondering if anyone has any } info on using an SEM while pregnant? I used the one here at SUNY } Potsdam during my first and third trimesters, as well as during the baby's } first 5 months. I keep having these awful scenarios running through my } mind about the old "Incredible Hulk" show where the guy was exposed to } gamma rays and became the Hulk!
I would like to embedd a loose piece of fabric for x-section analysis. The x-section face would be about 10mm wide. I am looking for a "soft" embedding material that can be sectioned with a razor blade. A low viscosity material is advantageous. Perhaps a silicone? Anything commercially available?
Any suggestions?
Thanks.
David Rose W.L. Gore & Associates 297 Blue Ball Road Elkton, MD 21921 410-506-2958
A large amount of information about the technical, economic and cultural developments associated with archaeological finds can be obtained from the usage marks on them. Such finds include domestic utensils and farm implements, weapons, jewellery and religious artefacts. Destruction of the finds is out of the question, and so scanning electron microscopy of the entire object has been an exception up to now.
We are a manufacturer of large chamber SEMs. With a volume of 2 cubic meters and aload capacity up to 300 kilograms most archeological artefacts can fit into our SEM.
Greetings from Germany Martin Klein ---------------------------------------------------------------------------- --- VisiTec Microtechnik GmbH Karl-Marx-Str. 14 D-23936 Grevesmuehlen/Germany Fon: +49-3881-79049 Fax: +49-3881-79048
----- Original Message ----- } From: tschwach {tschwach-at-mindspring.com} To: Microscopy ListServer {} Sent: Tuesday, October 05, 1999 9:38 PM
Back in the dark ages, before the invention of computers & scanners, "intensifier" solutions were used to darken black & white negatives that were too light. They may still be available in large photo shops. I would talk to someone experienced in the dark room. If you decide to darken the negative chemically, first try it on one or two frames.
Aydin
On Tue Oct 05 20:35:19 1999, "Donald Delaney" {delaneyd-at-mcw.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Here is my standard quick protocol which works 90% of the time.
Fix 30min. wash pbs buffer 2 x 5min ea. post fix 4% osmium tetroxide 5 min (fume hood!!! a real nasty) wash 2 x 5 min dH2O 50,75,95,100,100% ethanols series 5 min each step Hexamethyldisilazane 2 x 5 min. (fume hood) Air dry mount coat (residule HMDS will really gunk up a sputtercoater.
if you really want to avoid dislodging or dissolving crystals, vapor fix with osmium tetroxide 2-3 days in the refridge and slowly air dry. Let me know if you have other questions.
You can also check the Tips & Tricks archives at :
http://www.biotech.ufl.edu/~emcl/
If you cannot post fix in the osmium, double all times. The osmium really toughens them up.
At 02:06 PM 10/5/1999 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I would like to add to Donna's information request. I need to know how these usage fees are used. To what extent are funds used to maintain equipment as opposed to pay technical support salaries. Roughly what level of support is needed from the institution or does anyone out there actually run a facility independently from usage fees. Thanks- Dave
Home of the 1999 NCAA Basketball National Champion HUSKIES !!! ************************************************************
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax)
The fees that we recover are only about 20-25% of our total budget. (Incl. salaries fringe etc.) The rest is supported from the University under a legislative mandate for our biotechnology program. We are very lucky. We use those funds mostly for supplies and student help. Right now there is me directing the lab but not doing much of the work, since I have taken on other duties. Three full time techs and one student helper do the bulk of the work.
Greg Erdos.
At 08:04 AM 10/6/99 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
Hi evrybody I'm sorry in my last mail I forgot to mention it was for a FEG SEM with thermal cathode In order to plan a budget for a future FEG SEM I would like to know the following point concerning the commercial thermal FEG.
-type of instrument -How long is the life time? -What is necessary to exchange, tip only or gun set? -How much it cost? -How long it takes to replace -which garantee on lifetime? -comments?
Please answer directly to author. Thank you very much for your help.
Las Year in the ICEM-14 in Cancun, Mexico, there were about three free works related with Analytical SEM of archeological samples. Some of this them from Mexico, this are som titles: =20 "Applications of environmental sem to the art preseervation and archeological studies", J. Arenas, J.L. Galv=E1n and M. Jose Yacaman. ININ, Mexico. "Microstructural studies paints from the totonaca civilization in Tajin" D. Mendoza-Anaya.....and Jose Yacaman.
The Abstracts are in Vol II or III (I just have I and IV), One of the Scientist who works with archaeological samples is Dr. Jose Yacaman from Institiuto Nacional de Investigaciones Nuecleares, address: Amsterdam No. 46-202. Col. Hipodromo Condesa, 060100 Mexico, D.F., Mexico. Fax (5)= 3297299
Dr. Yacaman, besides, was the chairman of local organizing Commnitee, and editor of the abstracts.
Is there anyone out there that uses large quantities of electron image = film? If so, what kind of dessicant or pre-pump procedure do you use to dessi= cate film quickly? We go through quite a bit of film (Kodak ISO-163 for EM) and are lookin= g for an 'evironmentally friendly' film dessicant. Currently, we are using P= 2O5 powder in a vessel that we place in our cylindrical vacuum pump, in whi= ch we dessicate our film prior to loading the cassette into the electron microscope. I've ordered recyclable dessicant in a canister to try out= , but wanted to see how others are dealing with this aspect of microscopy.
Look forward to hearing from you all :)
Figen Seiler, Microscopist Abbott Labortories Department of Microscopy & Microanalysis
There is some new software, called "Lucis" which is really great for this purpose. Suggest that you give them a shout at Image Content Technologies, 860-223-4710 or wwww.imagecontent.com. Ron Lund is VP sales.
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 08:49 AM 10/5/99 -0500, Donald Delaney wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You are going to laugh, but I have used both a cork and a carrot for this exercise with fabric and got cross sections good enough to hold up in court. In either case, cut the "supporting material" lengthwise, place a small piece of fabric on the cut, replace the upper section and wind firmly with sewing thread. Cut with razor blade.
Best of luck! Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 06:56 AM 10/6/99 -0400, drose-at-wlgore.com"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Don: I have had great success with my Agfa Duoscan with light negatives by adjusting the gamma to a level of about 1.0 and then adjusting the manual TFS by pulling the slidebar to the left to adjust the sensitivity of the scan (These adjustments are done in the scanner software and not in Photoshop). Your scanner software might be different, but it should have similar scan sensitivity settings. If you need further help, please call me at 817 272-5496. Good luck, Mike Coviello Lab Manager UT Arlington
Donald Delaney wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have some negatives taken on the TEM which developed extremely light. I } was wondering if anyone knows any tricks on photoshop for generating good } quality prints. I typically adjust the levels and condense the size of the } picture, but I am still not completely happy about the quality of the prints. } } Don Delaney
All else being equal, I'd rather expose and process film for optimum density and contrast --neither under nor overexposing. That doesn't always work out in practice, and you can't always reshoot (or even find) a given sample area in TEM. (The question was: how to deal with underexposed film).
The reason for scanning TEM negatives as positive transparencies is that the control software that comes with ordinary flatbed scanners (such as mine) automatically applies a lookup table to negative scans to correct for the low contrast of 35 mm negative film. The result from a negative scan of a contrasty TEM negative is posterization --stairstepping of contrast levels on the output image. For reasons known only to the manufacturers, they output positive scans without applying the correction. (I've commented on this to the listserver before).
Some experienced TEM darkroom practitioners feel that they get better control over tonal adjustments by printing and scanning the prints. I'm not sure I want to defend this because it's not my personal practice. However, I've seen some pretty nasty film faults recovered in the darkroom.
Larry -- From: Dr. Gary Gaugler Sent: Tuesday, October 5, 1999 5:38 PM To: MSA listserver Subject: RE: Photoshop Negative Processing
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
The main problem is that if the information is not in the neg, it cannot be created using Photoshop. It can be subtracted or contrast enhanced but not created. It is always better to overexpose whenever deciding which way to go.
Also, scanners tend to produce better results when set to negative rather than positive. This is because the neg has an inherently greater tonal range than a positive and the scanner tries to capture that. If you have a neg, scan as a neg. I have always achieved better results this way.
Printing an underexposed neg is going to produce a contrasty print. Still no additional info. Actually less. The only viable method of fixing an underexposed neg is to re-shoot it.
gary g.
At 03:15 PM 10/5/99 , you wrote:
} Photoshop processing can help, but the place to start is in scanning. If you } have a 30-bit or better yet a 36-bit (color) scanner, you should be able to } manipulate the scanner's tone controls (black, white, and gamma) to optimize the } scanner's 8-bit (grayscale) output. You might also try superimposing a neutral } density filter on the film negative to match the work density to the scanner's } range. The filter can be another piece of light-exposed and developed TEM } film. It just needs to have a uniform (gray) background. Always scan TEM } negatives as positive transparencies; then invert the contrast in the scanner or } Photoshop. } } A work-around, especially if you don't have a good scanner, is to print the } negative in the darkroom to get the contrast and density you want and then scan } the print. } } To further adjust the tonal quality in Photoshop, use Levels or Curves } adjustment layers rather than applying these commands directly. This will avoid } much of the data loss in applying multiple corrections. Use the Multiply } blending mode to increase the contrast from a light negative, and apply as many } layers as needed to get the contrast and background brightness right for your } printer. If you use unsharp mask filtering (highly recommended after scanning) } and background leveling, apply these to layers before the tonal adjustments. } Learning to use Layers in Photoshop takes some time, but is highly worthwhile. } Good luck. } } Larry Thomas } } Larry Thomas } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto: Larry.Thomas-at-pnl.gov } } } } } From: Donald Delaney } Sent: Tuesday, October 5, 1999 6:49 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Photoshop Negative Processing } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have some negatives taken on the TEM which developed extremely light. } I } was wondering if anyone knows any tricks on photoshop for generating } good } quality prints. I typically adjust the levels and condense the size of } the } picture, but I am still not completely happy about the quality of the } prints. } } Don Delaney } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Please contact me if you have an old Hitachi HU11E or HU125E that is available for parts. I am most interested in vacuum gauges and associated electronics.
I teach a TEM/SEM course at a private liberal arts college and operate a HU125E without a service contract and I am in need of spare parts!
Thanks
Robert
Robert Fitton Luther College Department of Biology 700 College Drive Decorah, IA 52101
Voice 319-387-1559 FAX 319-387-1080
Enjoy a visit to our website: http://www.luther.edu/dept/bio.htm
Folks, Regarding the offer of free ISI model 40 parts: Bill Blackburn whishes to thank the many people who have responded to the posting. All the parts have now been given out. Bill regrets he didn't have enough to satisfy all who contacted him. ####################################################### Original Message: "Microscopy Listserve Members--- I have 20 new filaments; several Wehnelt assemblies; SEI, TE, CL?, detectors; Scintillator disks; and other small items from an ISI Model 40 SEM. They are free if someone can use them. Please get in touch with me directly via email at probe-at-mailbox.cc.binghamton.edu or phone 607 777-2832."
Best Regards, Bill Blackburn Geology Department Binghamton University Binghamton, NY 13902-6000
Anyone have any experience or references to pass along??
} From: Keith Peacher {KPeacher-at-embrex.com} } To: sdw-at-biotech.ufl.edu } Subject: Freezing Tissue } Date: Wed, 6 Oct 1999 12:58:08 -0400 } X-Mailer: Internet Mail Service (5.5.2448.0) } } Hi Wiz, I am looking to freeze whole chicken eggs at different embryonic stages. I want to preserve tissues for further analysis. I was searching for ideas when I came across your web site. I thought I would ask you if you could provive some insight on getting started with this task. } A. Keith Peacher } Research Associate II } {mailto:kpeacher-at-embrex.com} Embrex, Inc. } P.O. Box 13989 } Research Triangle Park, NC 27709-3989 Tel.(919)-941-5185 } Fax.(919)-941-5186 } {http:\\www.embrex.com} }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I am trying to repair a cold cathode luminescence stage from a company called Technosyn. The system was bought from S&M Microscopes, Inc. in Colorado Springs, Colorado. I have not been able to contact S&M with the phone numbers I have, or locate any electronic drawings of the system. My best guess is that the E-gun or high voltage supply has failed. Any contact information or source of drawings from any of you on the list would be very helpful.
Thanks Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Try this. Duplicate the image and then paste the dup'd image to the original. Use multiply for the overlay mode 100%. Alternatively, you can just use Image-ApplyImage. Use the same image as the source and target and use 100% opacity and multiply for blending. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Donald Delaney To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
I have some negatives taken on the TEM which developed extremely light. I was wondering if anyone knows any tricks on photoshop for generating good quality prints. I typically adjust the levels and condense the size of the picture, but I am still not completely happy about the quality of the prints.
We have some non-linear filters and other manipulation facilities in our software, analySIS. If you are willing to part with one of your negatives for a while or if you can send us a scanned or otherwise digitized image, I can try and see if we can do something for you.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
At 08:49 AM 10/5/99 -0500, Donald Delaney wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have experience with (or references for) immunogold labeling of cultured cells for ultrastructural analysis? Pre-embedment is preferred over post-embedment methodology. I am trying to preserve polyribosomes concomitant with 1.4nm gold immunolabeling of cytoplasmic proteins. 0.1% saponin and/or triton permeabilization following mild aldehyde fixation enhances antibody penetration, but compromises ultrastructural preservation.
Is there a less destructive way to obtain sufficient antibody penetration into monolayer cultures, or must I look at brain slices?
Don't jump to conclusions here: I have friends (scientists) who have absolutely nothing to do with either SEM or TEM and their teenagers show the same symptoms. Perhaps the X-rays are distributed through the ventilation system?? Or along Power lines?? My personal favorite: a government conspiracy!!
But seriously: If I were pregnant, I would have the SEM checked out with a radiation meter, just to make sure there are no modifications on the instrument that could cause a radiation leak. Since SEMs usually have low acceleration voltages and solid specimen chambers, I would, however, NOT expect to find anything. It's really more for peace of mind.
TEMs, on the other hand, is something that can potentially be more dangerous. Not only is the acceleration voltage much higher (several hundred keV or more), but the radiation is also produced fairly close to where you don't want to have it. In addition, many TEMs have axially mounted cameras, which need to be shielded also. A radiation test should be done on a regular basis anyway, because a leaky TEM is not something you want to work on.
If you measure the radiation, there are limits set by OSHA concerning radiation exposure for differnt types of exposure.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: tschwach[SMTP:TSCHWACH-at-MINDSPRING.COM] } Sent: Wednesday, October 06, 1999 12:22:41 PM } To: Scan Service; Randy & Jenna & Orin Brown } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: pregnancy and the SEM } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thank you for explaining what is apparently "wrong" with my teenage boys. All this time I thought it was the embedding chemicals I used for TEM.
} It's obviously a little late to be asking about this, since the baby } is fine and almost 10 months old... BUT I was wondering if anyone has any } info on using an SEM while pregnant? I used the one here at SUNY } Potsdam during my first and third trimesters, as well as during the baby's } first 5 months. I keep having these awful scenarios running through my } mind about the old "Incredible Hulk" show where the guy was exposed to } gamma rays and became the Hulk!
Jenna -
There's no radiation hazard from any modern SEM that has no "non-stock" column modifications. There ARE chemical hazards associated with specimen prep.
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
David, I get better "cross sections" of polymer films/fibers/fabrics by sealing the sample in parafilm, freezing it on a copper block sitting in LN and fracturing it with a frozen razor blade. I use a hemostat to clamp a single edge razor blade. I cut down a styrofoam container so that it is about 4" deep. If you don't have LN, dry ice also works. I freeze the sample, cool the blade, fracture, retrieve the pieces discarding the parafilm and check themunder a dissecting scope at 45X. Under reflected light, the fracture face is shiny compared to the edge cut with scissors or a razor blade. I mount the piece on edge/frac. face-up using double sticky carbon tabs. Usually the pieces are small enough that they stand up. If they are flimsy like nylon or polycarbonate filters, I place a piece of capillary tube onto the first double sticky tab and cover it with a second double sticky tab and lean the piece against the raised support---you'll be able to place several pieces along each side of the tube. Follow with C evaporation. Imaging and X-ray analysis are routine although it may be necessary to tilt the sample's off from the 30 tilt generally used for the take-off angle. I like to use line-profile analysis to demonstrate surface modifications of polymer films. Best of luck! Rosemary
-----Original Message----- } From: Thomas, Larry {Larry.Thomas-at-pnl.gov} } All else being equal, I'd rather expose and process film for optimum density and } contrast --neither under nor overexposing. That doesn't always work out in } practice, and you can't always reshoot (or even find) a given sample area in } TEM. (The question was: how to deal with underexposed film). ============= Getting it right is sure the best. But with the ability to combine images you can get greater detail in 3 negitives one normal, one under and one over exposed. } } The reason for scanning TEM negatives as positive transparencies is that the } control software that comes with ordinary flatbed scanners (such as mine) } automatically applies a lookup table to negative scans to correct for the low } contrast of 35 mm negative film. The result from a negative scan of a contrasty } TEM negative is posterization --stairstepping of contrast levels on the output } image. For reasons known only to the manufacturers, they output positive scans } without applying the correction. (I've commented on this to the listserver } before). } } Some experienced TEM darkroom practitioners feel that they get better control } over tonal adjustments by printing and scanning the prints. I'm not sure I want } to defend this because it's not my personal practice. However, I've seen some } pretty nasty film faults recovered in the darkroom.
I have recovered some of the those nasty faults in the dark room. I promise you you can do more with photo shop than with an enlarger. I took images I made 30 years ago that were unprintable and made decent looking digital prints. There was not one bit more of detail there but I could stretch the contrast range until it looked OK.
Every time you process the image you loose information. It never gets better than the original. You may be able to get different data from the original but never more. Any gain in one area results in a loss in another.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK 405 624-2855 GMT -6:00 www.couger.com/gcouger
Figen: I don't know about the emulsion thickness of the ISO-163, but the SO film has a thicker emulsion than the 4489 and so it holds a bit more water.
P2O5 other than con H2SO4 (clearly not suitable) is about the most powerful desiccant. So if you care for very dry film that is the best to use for a final desiccation step (dogma?!). There are other means which help to make the expensive and potentially dangerous P2O5 go much further or for some instruments even redundant. Note that adding liquid water to P2O5 produces phosgene, a rather nasty gas.
With a lot of film in stock, the internal plastic bags could be cut open and the film stored (light-tight of course) in the fridge or freezer. Fridge and freezer desiccate, it just takes a while. When removing, enclose the container in a plastic bag until it reaches room temperature.
Desiccation under vacuum is usually combined with P2O5, however, if the vacuum chamber is at a somewhat elevated temperature (37 degrees overnight), many people would find that degree of desiccation sufficient.
Silica Gel is not very powerful but would help in the process when added to the vacuum desiccator. I have not tried to use silica gel with bulk film in a (non-vacuum) desiccator. I expect if the film was stored in vented boxes, that within a month the film moisture level would drop to a small percentage of fresh film. I think that this may be the simplest means of pre-desiccating large amounts of film.
As noted, opening the film envelopes before refrigeration and soon after the film is received, does help the drying process. It is much more effective if the film could be removed from the envelopes. Ideally the film would then be placed in "vented" boxes with a light trap. These boxes could be used in the fridge/ freezer or a desiccating cabinet during the first stage of desiccation; for the final stage, perhaps with desiccant, the film must be in sheetfilm holders.
Microscopy aside and in the name of conservation: collect the liquid P2O4 (now phosphoric acid) soon somebody will want some to paint over a rusty surface, which is then followed by an undercoat. It's the best rust inhibitor (dogma!?) and saves some phosphorus from polluting the waterways. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, October 07, 1999 1:19 AM, Seiler,Figen [SMTP:figen.a.seiler-at-abbott.com] wrote:
} Is there anyone out there that uses large quantities of electron image film? } If so, what kind of dessicant or pre-pump procedure do you use to dessicate } film quickly? } We go through quite a bit of film (Kodak ISO-163 for EM) and are looking for } an 'evironmentally friendly' film dessicant. Currently, we are using P2O5 } powder in a vessel that we place in our cylindrical vacuum pump, in which we } dessicate our film prior to loading the cassette into the electron } microscope. I've ordered recyclable dessicant in a canister to try out, but } wanted to see how others are dealing with this aspect of microscopy. } } Look forward to hearing from you all :) } } Figen Seiler, Microscopist } Abbott Labortories } Department of Microscopy & Microanalysis } } E-mail: figen.a.seiler-at-abbott.com
When I worked in labs that had radiation hazards I made checks on radition leaks every week for my own satisfaction. I don't trust anybody to do it for me when grad students are involved.
It is also amazing what you find is hot.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK 405 624-2855 GMT -6:00 www.couger.com/gcouger } } TEMs, on the other hand, is something that can potentially be more } dangerous. Not only is the acceleration voltage much higher (several } hundred keV or more), but the radiation is also produced fairly close to } where you don't want to have it. In addition, many TEMs have axially } mounted cameras, which need to be shielded also. A radiation test should } be done on a regular basis anyway, because a leaky TEM is not something } you want to work on. } } If you measure the radiation, there are limits set by OSHA concerning } radiation exposure for differnt types of exposure. } } Michael } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } ---------- } } From: tschwach[SMTP:TSCHWACH-at-MINDSPRING.COM] } } Sent: Wednesday, October 06, 1999 12:22:41 PM } } To: Scan Service; Randy & Jenna & Orin Brown } } Cc: Microscopy-at-sparc5.microscopy.com } } Subject: Re: pregnancy and the SEM } } Auto forwarded by a Rule } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Jim Phosgene is the acid chloride of carbonic acid, otherwise known as carbonyl chloride, COCl2. You would need to be an alchemist to make it from phosphorous pentoxide and water. If you can do it, I would like to consult you about a little gold production project I have in mind! Chris } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Figen: } I don't know about the emulsion thickness of the ISO-163, but the SO film has a } thicker emulsion than the 4489 and so it holds a bit more water. } } P2O5 other than con H2SO4 (clearly not suitable) is about the most powerful } desiccant. So if you care for very dry film that is the best to use for a final } desiccation step (dogma?!). There are other means which help to make the } expensive and potentially dangerous P2O5 go much further or for some } instruments even redundant. Note that adding liquid water to P2O5 produces } phosgene, a rather nasty gas. } } With a lot of film in stock, the internal plastic bags could be cut open and } the film stored (light-tight of course) in the fridge or freezer. Fridge and } freezer desiccate, it just takes a while. When removing, enclose the container } in a plastic bag until it reaches room temperature. } } Desiccation under vacuum is usually combined with P2O5, however, if the vacuum } chamber is at a somewhat elevated temperature (37 degrees overnight), many } people would find that degree of desiccation sufficient. } } Silica Gel is not very powerful but would help in the process when added to the } vacuum desiccator. I have not tried to use silica gel with bulk film in a } (non-vacuum) desiccator. I expect if the film was stored in vented boxes, that } within a month the film moisture level would drop to a small percentage of } fresh film. I think that this may be the simplest means of pre-desiccating } large amounts of film. } } As noted, opening the film envelopes before refrigeration and soon after the } film is received, does help the drying process. It is much more effective if } the film could be removed from the envelopes. Ideally the film would then be } placed in "vented" boxes with a light trap. These boxes could be used in the } fridge/ freezer or a desiccating cabinet during the first stage of desiccation; } for the final stage, perhaps with desiccant, the film must be in sheetfilm } holders. } } Microscopy aside and in the name of conservation: collect the liquid P2O4 (now } phosphoric acid) soon somebody will want some to paint over a rusty surface, } which is then followed by an undercoat. It's the best rust inhibitor (dogma!?) } and saves some phosphorus from polluting the waterways. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Thursday, October 07, 1999 1:19 AM, Seiler,Figen } [SMTP:figen.a.seiler-at-abbott.com] wrote: } } } Is there anyone out there that uses large quantities of electron image film? } } If so, what kind of dessicant or pre-pump procedure do you use to dessicate } } film quickly? } } We go through quite a bit of film (Kodak ISO-163 for EM) and are looking for } } an 'evironmentally friendly' film dessicant. Currently, we are using P2O5 } } powder in a vessel that we place in our cylindrical vacuum pump, in which we } } dessicate our film prior to loading the cassette into the electron } } microscope. I've ordered recyclable dessicant in a canister to try out, but } } wanted to see how others are dealing with this aspect of microscopy. } } } } Look forward to hearing from you all :) } } } } Figen Seiler, Microscopist } } Abbott Labortories } } Department of Microscopy & Microanalysis } } } } E-mail: figen.a.seiler-at-abbott.com } }
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
Assuming that antigens have been adequately preserved, the potential success of pre-embedding immunogold labeling seems to depend on the hydrodynamic size of the reagents, the degree to which membranes are permeabilized and the extent of cross-linking of the cytoplasm. And last but not least: time and temperature of incubation. It is advisable to use the smallest possible reagents, for instance single Fab fragments instead of complete IgG. Penetration can be achieved with detergents, but their application affects the way the cellular organization is preserved. A milder treatment uses sodium borohydride, used in immunofluorescence to reduce autofluorescence. The way it works as a substance that enhances penetration has, to my knowledge, never been elucidated, but used at a concentration of 0.1% in PBS for something like 15 minutes it opens up structures and allows gold particles to enter the cytoplasm. This way we obtained highly efficient tubulin and actin labeling in cultured PTK2 cells, fixed in 0.5% glutaraldehyde for 15-30 minutes. Likewise Van Lookeren Campagne obtained labeling of B-50 protein and MAP2 in cultured neuron cells. In our experience and that of several pioneers in pre-embedding immunogold technology the degree of cross-linking by glutaraldehyde fixation especially seems to be a more serious (and often underestimated) issue to deal with. The higher the degree of cross-linking, the longer distances reagents have to travel to find their targets. The solution? Be patient, give it time for the reagents to find their way to the target molecules by diffusion. And one last thing: if it takes a while for antibodies and reagents to get in, it will also take a while for unbound reagents to get washed out again after the incubation steps, so washing steps have to be substantially longer than in postembedding.
Good luck, Jan =========================== Jan Leunissen AURION http://www.aurion.nl Costerweg 5 6702 AA Wageningen phone: (31)-317-497676 fax: (31)-317-415955 You will find more tech info on our website.
While the subject of thermally assisted FESEMs is on your minds (Serge's note), we have been checking into who still offers these beasts in their product line. We know of Leo, and FEI. Are there any other manufacturers that have not gone completely into the cold for their lab/analytical SEMs?
On Wed, 6 Oct 1999 drose-at-wlgore.com-at-Sparc5.Microscopy.Com wrote:
} I would like to embedd a loose piece of fabric for x-section analysis. The } x-section face would be about 10mm wide. I am looking for a "soft" embedding } material that can be sectioned with a razor blade. A low viscosity material is } advantageous. Perhaps a silicone? Anything commercially available?
Perhaps you could infiltrate the fabric with molten paraffin wax at 60^C under vacuum, and then cut as one normally does cut a section of plant tissue or whatever. If even 60^C temperature is a problem, then there are several solvents with a high freezing point you could use, together with dry ice cooling.
+------------------------------------------------------------------------+ | 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 position is for an SEM technician at Kopin Corporation, the world=92= s leading provider of advanced HBT transistor wafers, technical design an= d manufacturing support services. The SEM technician will be responsible = for preparing and analyzing SEM samples. These will primarily consist of Ga= As based semiconductor devices and calibration structures. The technician = will also be trained in and expected to perform additional structural and electronic measurements as needed. These include Polaron (electrochemic= al etching with C-V profiling), Hall measurements, photoluminescence, x-ra= y diffraction and photoreflectance. The technician must have the ability = to handle parallel tasks effectively, solve minor problems in the lab, and=
maintain a clean, organized, and safe working environment. Good communication and interpersonal skills are essential. The ideal candida= te will have a 2-year college or technician degree in a science or enginee= ring field and no less than 2 years SEM experience. Familiarity with semiconductor process technology is a plus. Kopin is an equal opportun= ity employer. Resumes accepted by fax, e-mail or mail. Interested candidate= s should contact:
Cheryl Messier Human Resources Kopin Corporation 695 Myles Standish Blvd. Taunton, MA 02780 Fax (508)824-6958 cheryl_messier-at-kopin.com =
The position is for an SEM technician at Kopin Corporation, the world=92= s leading provider of advanced HBT transistor wafers, technical design an= d manufacturing support services. The SEM technician will be responsible = for preparing and analyzing SEM samples. These will primarily consist of Ga= As based semiconductor devices and calibration structures. The technician = will also be trained in and expected to perform additional structural and electronic measurements as needed. These include Polaron (electrochemic= al etching with C-V profiling), Hall measurements, photoluminescence, x-ra= y diffraction and photoreflectance. The technician must have the ability = to handle parallel tasks effectively, solve minor problems in the lab, and=
maintain a clean, organized, and safe working environment. Good communication and interpersonal skills are essential. The ideal candida= te will have a 2-year college or technician degree in a science or enginee= ring field and no less than 2 years SEM experience. Familiarity with semiconductor process technology is a plus. Kopin is an equal opportun= ity employer. Resumes accepted by fax, e-mail or mail. Interested candidate= s should contact:
Cheryl Messier Human Resources Kopin Corporation 695 Myles Standish Blvd. Taunton, MA 02780 Fax (508)824-6958 cheryl_messier-at-kopin.com =
Yes The Camscan MX2540 SF thermal FESEM springs to mind. Read all about it, and the advantages of thermal FE on their website: http://camscan.co.uk/index.htm
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } While the subject of thermally assisted FESEMs is on your minds } (Serge's note), we have been checking into who still offers these } beasts in their product line. We know of Leo, and FEI. Are there } any other manufacturers that have not gone completely into the } cold for their lab/analytical SEMs? } } Thank you, } Darrell Miles } } }
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
Dear Listers, Those people concerned with pregnancy and SEM should remember that the SEM uses the same accelerating voltage (or lower) as a TV, without the potential leakage from a glass front screen. TEMs should be carefully screened for leakage. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
On the tread of FE-SEM emitters I would be very interested in hearing what people have to say about the differences between cold field emitters and Schottky emitters. I know what the manufacturers say but what about expert users? Thanks for any input.
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
Does anyone know how to make those clear plastic blocks with embedded specimens? Sometimes you see them with flowers or bugs inside used as paperweights. We would like to make some with some special stuff inside for display in our lab.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
On the tread of FE-SEM emitters I would be very interested in hearing what people have to say about the differences between cold field emitters and Schottky emitters. I know what the manufacturers say but what about expert users? Thanks for any input.
------------------------------ Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
} } Jim } Phosgene is the acid chloride of carbonic acid, otherwise known as } carbonyl chloride, COCl2.
Jim was probably trying to type phosphine which is produced by the reaction of water and elemental phosphorous. If there is concern of producing phosphine from water and phosphorous pentoxide, my guess is it would be from unreacted P in the P2O5.
In terms of dessication, it would seem that a simpler and more environmentally friendly method would be a nitrogen purged glove box. Has anybody compared relative pump down times of vacuum dessicated film boxes versus dry nitrogen purged?
Of course everybody knows that MOXTEK makes ultrathin windows for Si(Li) detectors, so I might be a little biased.
These windows are very fragile, but have a surprisingly long lifetime. We still have windows out in the field that we made 9 years ago. In the early days we found that some microscope models had problems during the venting that would cause particles to impact the window film, piercing it, but these bugs have been mostly worked out.
The EDS manufacturers all have better data on failure rates than we do, since we rarely hear from a microscopist about his experience good or bad. I would suggest asking them for this data if you are concerned.
Based on our knowledge, I can say that there does not seem to be a "wear out" mechanism, and that a good window will be good indefinately. The biggest cause of window failure in the ones we have seen back from the field is that the window has been touched.
best regards mark
Mark W. Lund, PhD VP Engineering } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"This is a YOUNG business...How can I tell you what YOUR job is when I don't know what MINE is?" --Pogo
Richie Sims wrote:
Hi, Everyone
I'm shortly going to buy a new EDS detector. I'd quite like the performance advantages of a 1-atmosphere UTW, but I'm concerned about the durability and longevity compared with standard Be windows.
Anyone out there got any relevant experience/views?
Either to the list, or directly to me, please.
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
We have one of each. Hands down the cold cathode instrument is the superior imaging tool but that instrument is optimized for hi-res imaging. Our hot cathode instrument is a analytical platform with poor vacuum / high vacuum modes. I believe a local space agency bought the next generation of our cold cathode SEM after a demo on ours. They were using a hot cathode instrument previously. Interestingly we bought a similar hot cathode instrument later but only because that is how the system came. It was purchased for it's poor vacuum capability.
I use the cold cathode instrument for all (imaging) but the lowest mag work ( {2k). It suffers from spherical aberration at lower mags. This is my SEM of choice. I use the hot cathode instrument for low mag work ( {2k), EDS or wet work. It has a larger area of view. At times I have used it up to x10K but only because I was there & had a favorable specimen.
The contrast & image detail of the cold cathode system is decidedly superior. I can also point out that the images can be acquired at 5 kV, some times less. I always use 30 kV for the hot cathode SEM.
Let's be clear....There are differences in the optics & secondary detection systems which account for some of the difference in image quality. Both instruments have a place.
On reliability, the cold cathode went on line in 1994. The instrument is anvil reliable. The hot cathode has been changed 2-3 times since 1995. This may have been a production yield problem. The current one has been in a while.
I have purposely avoided using brand names & model #s because people tend to associate properties or opinions of individual instruments with entire product lines. If you would like specifics, please contact me off line.
Meanwhile if your looking to buy, I suggest that you have vendors image a variety of materials before you make a firm deci$sion. Also conditions of final payment should include being able to replicate those images on you instrument.
Bruce Brinson Rice U.
Stephen McCartney wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } On the tread of FE-SEM emitters I would be very interested in hearing what } people have to say about the differences between cold field emitters and } Schottky emitters. I know what the manufacturers say but what about expert } users? Thanks for any input. } } ------------------------------ } Stephen McCartney } Research Associate } Virginia Tech } Materials Institute } 2108 Hahn Hall } Blacksburg, VA 24061-0344 } USA } } TEL: 540-231-9765 } FAX: 540-231-8517 } ------------------------------
would like to receive sugestions about etching of gray cast irons, I want to find how to reveal eutectic cells and correlation them with machining. thanks.
Alonso de la Garza S. Metallograpy laboratory. Facultad de Ingenieria,UASLP
- } } } On Wed, 6 Oct 1999 drose-at-wlgore.com-at-Sparc5.Microscopy.Com wrote: } } } I would like to embedd a loose piece of fabric for x-section analysis. The } } x-section face would be about 10mm wide. I am looking for a "soft" embedding } } material that can be sectioned with a razor blade. A low viscosity material is } } advantageous. Perhaps a silicone? Anything commercially available? } } Perhaps you could infiltrate the fabric with molten paraffin wax at 60^C } under vacuum, and then cut as one normally does cut a section of plant } tissue or whatever. If even 60^C temperature is a problem, then there are } several solvents with a high freezing point you could use, together with } dry ice cooling. }
If cold is a problem benzene freezes as 5 c. I have no idea how it cuts?
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK 405 624-2855 GMT -6:00 www.couger.com/gcouger
There was a very interesting and controverse discussion of this toppic on the list back in February 1997. You should check the archive of the listserver:
At 15:39 07.10.99 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
There is a forthcoming conference in Toledo, Spain on Field emission in general. There will be quite a lot of discussion on various types of emission, from manufacturers, Academic labs and end users. More info is at www.cmp-cientifica.com.eurofe
Based on our experience, the Schottky tips are less trouble as they don't require flashing every few hours to keep them in good condition. I had a comparison from Philips Electron Optics on this, I'll try to dig it out if anyone is interested.
Tim
++++++++++ On the tread of FE-SEM emitters I would be very interested in hearing what people have to say about the differences between cold field emitters and Schottky emitters. I know what the manufacturers say but what about expert users? Thanks for any input.
Dear All, I enclose details of a post-doc vacancy at the University of Barcelona, Spain. The starting date would be about April-May 2000, so we have extended the deadline for applications. Any one interested please reply directly to paqui-at-el.ub.es and/or send applications and a CV by mail before 30 November 1999.
Kind regards
F. Peir=F3
**************************************************************************= ** Laboratory: Electronic Materials and Engineering, Department of Electronics, University of Barcelona.
Duration: 12-18 months, starting April-May 1999.
Jon,
You can try Castolite resin. It is fairly inexpensive if purchased from US Plastics (check their web site) and it is very clear. You just need a mold to hold the shape. I'm not sure if that is what everyone else uses but it should work out OK.
______________________________________________ Roberto Garcia Senior Analyst, Metallography NC State University / Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh, NC 27695-7531 (919) 515-8628 (919) 515-6965 Fax rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif ____________________________________________
----- Original Message ----- } From: Jon Krupp {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 07, 1999 4:21 PM
Alonso -
We use a recipe we were kindly donated by a gray iron foundry in Wisconsin. They call it "Stead's etch" but it is slightly different from the "Stead's etch" I find in my metallography bible - (Vander Voort - Metallography Principles and Practice - no longer in print). The foundry did a good bit of work to figure out what worked consistently and found that it was critical to use denatured alcohol - pure grain alcohol is the best. I've found that immersing a polished sample in a fresh batch of this works well. It's worked on almost all the gray irons I receive from different foundries.
Recipe 1- 2 grams CuCl2-2H2O 8 grams MgCl2-6H2O 4 ml HCl 100 ml grain alcohol (etch time up to 2 minutes)
Recipe 2- 1 gram CuCl2-2H2O 4 grams MgCl2-6H2O 2 ml Hcl 100 ml grain alcohol (etch time up to 7-8 minutes)
Examine under standard lighting conditions in the optical microscope
If you over-etch the boundaries disappear. If you under-etch the boundaries are too thick and you miss small cells. You need to etch - check, etch - check......
Good luck and let me know if it doesn't work as Vander Voort has some other recipes.
Robin Griffin Materials and Mechanical Engineering The University of Alabama at Birmingham rgriffin-at-eng.uab.edu
-----Original Message----- } From: alonso de la garza san miguel [mailto:alonsod-at-uaslp.mx] Sent: Thursday, October 07, 1999 9:33 PM To: Microscopy-at-Sparc5.Microscopy.Com
would like to receive sugestions about etching of gray cast irons, I want to find how to reveal eutectic cells and correlation them with machining. thanks.
Alonso de la Garza S. Metallograpy laboratory. Facultad de Ingenieria,UASLP
Although I have not done it myself I know that Carolina Biological Supply sells the polyester embedding kits with instructions (Plastomounts) as well as already mounted specimens. They can be reached at 1-800-334-5551 or www.carolina.com.
Louie
At 1:21 PM -0700 10/7/99, Jon Krupp wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
Good ideas, but what effect will these solvents have on any polymeric component of the fabric?
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 08:40 AM 10/8/99 +0100, Robert H. Olley wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At 01:21 PM 10/7/1999 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
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} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi: } } Does anyone know how to make those clear plastic blocks with embedded } specimens? Sometimes you see them with flowers or bugs inside used as } paperweights. We would like to make some with some special stuff inside for } display in our lab. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
Hi All, I've had very good results with a material named CASTOLITE AP. It is a water clear resin when cured and hard enough to be optically polished if so desired. It is manufactured by the Castolite Co. Get their literature and choose the resin that best meets your needs. They provide good directions for the embedding of biological specimens.
Hello all, I am looking for a good example of simulated diffraction contrast images for a book. The sort of thing I have in mind is a series of two-beam experimental and simulated images of a dislocation or stacking fault; credit would of course be given in the caption accompanying the figure. Does anyone do this any more? Just about everyone I ask has high res. phase contrast (HREM) image simulations, but nobody does the low mag, diffraction contrast stuff. I would have thought that it would be relatively easy to do on a PC (if only I had the time to learn C++)...
We have an embedding material called PolyMet which is a polyester materia= l that cures crystal clear. It generally cures crystal clear although it must usually cure 6 to 8 hours (overnight is best) at room temperature an= d is not generally as hard as a standard metallurgical mount. We sell this=
} } } } } 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 Jon Krupp }
Hi:
Does anyone know how to make those clear plastic blocks with embedded specimens? Sometimes you see them with flowers or bugs inside used as paperweights. We would like to make some with some special stuff inside f= or display in our lab.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu {
Can anyone give me some refernces for a method of fixing tissue in liquid propane, then dehydration in acetone at -80C ? I think that is the sequence. Anyway, a friend was telling me about this method and now I cannot get in touch with him to find out more. Thanks Mary
I remember, that at one time as a child I had a "kit" where I could embed things in clear plastic (for sea shells, coins, and all sorts of other stuff). I don't know if these kits are still being sold. Perhaps a trip to the local toy shop might help.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Louie Kerr[SMTP:LKERR-at-MBL.EDU] } Sent: Friday, October 08, 1999 7:23:32 AM } To: jmkrupp-at-cats.ucsc.edu; Microscopy-at-sparc5.microscopy.com } Subject: Re: Plastic embedding for display } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Jon,
Although I have not done it myself I know that Carolina Biological Supply sells the polyester embedding kits with instructions (Plastomounts) as well as already mounted specimens. They can be reached at 1-800-334-5551 or www.carolina.com.
Louie
At 1:21 PM -0700 10/7/99, Jon Krupp wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have used this process with great success. It was introduced to me as "freeze substitution" and gives excellent preservation of tissues. The two ways that I have used liquid propane to do this is through either "jet freezing" or "plunge freezing". Jet freezing requires a specialized jet freezing apparatus, however an effective plunge freezing apparatus can be put together without to much difficulty, and I have found this method to give me superior results to the jet freezing.
I'm sure there are others on the list with more expertise than I, but if you would like to reply to me personally, I could tell you how to put together an effective plunge freezing apparatus without to much difficulty. I think this would be better than going through it all on the list.
Tim Wakefield ----- / 101 Cary Hall / | \ / Auburn University, AL / --|-- \/ 36849 \ | /\ 334-844-3908 \ | / \ ----- \
On Fri, 8 Oct 1999, Mary C. Pfauth wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone give me some refernces for a method of fixing tissue in liquid } propane, then dehydration in acetone at -80C ? I think that is the } sequence. Anyway, a friend was telling me about this method and now I } cannot get in touch with him to find out more. Thanks Mary } } John P.B. & Mary } mpfauth-at-teleport.com } } }
"Tim E. Harper" {tim-at-cmp-cientifica.com} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} www.cmp-cientifica.com.eurofe } } Based on our experience, the Schottky tips are less trouble as they don't } require flashing every few hours to keep them in good condition. I had a } comparison from Philips Electron Optics on this, I'll try to dig it out if
} anyone is interested. } } Tim } } ++++++++++ } On the tread of FE-SEM emitters I would be very interested in hearing what
} people have to say about the differences between cold field emitters and } Schottky emitters. I know what the manufacturers say but what about expert } users? Thanks for any input. } } ************ } Tim E. Harper CMP Cientifica s.l. } Nanofabrication & Advanced Materials Analysis Consultants } Apdo Correos 20, 28230 Las Rozas, Madrid, Spain } Tel: +34 91 640 71 85 Fax +34 91 640 71 86 } E-mail: mailto:Tim-at-cmp-cientifica.com } http://www.cmp-cientifica.com/ } } } } } } } } } ************************************************************************** **
Hitachi S-510 SEM+X-ray detector and image acquisition system for sale.
- The microscope is 13 years old, It has been under Hitachi service contract ever since it was bought and it is in excellent working condition. - X-ray detector (Canberra) has three modes: Be, UTW and windowless, for light element detection. It has undergone a comprehensive detector restoration ~8 months ago and is still under warranty. - Custom-built image acquisiton system with a Mitsubishi video printer.
The whole setup is on sale for $12,000. The buyer will pay for dismantling and shipping of the instrument. All interested parties should contact Prof. Yip-Wah Chung. e-mail: ywchung-at-nwu.edu phone: (847) 4913112
Thanks, Murat Guruz ___________________________________________ Murat U. Guruz Dept. of Materials Science and Engineering Northwestern University Ph (847) 491-3216 491-7798 Fax (847) 491-7820 m-guruz-at-nwu.edu http://vpd.ms.nwu.edu/vpdgroup/mg.htm
During Microscopy and Microanalysis '99 in Portland many of you donated= sand to the Project Micro sand collection. I would like to thank all who donated. Thanks to your donations we now have sand from every continen= t on our planet. Some of the sands were very interesting. However, many donations just appeared at the project micro booth with no names on the= m. If you were one of our anonymous donors and would like to get credit for y= our donation, please e-mail me at the address below. We generally include = the name and affiliation of the donors when we send sand to teachers and microscopists. If you would like to make a donation please mail your s= and to the address below. If you would like to request some sand for educatio= nal purposes, contact me or read my next e-mail message to the list.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com =
Project Micro would like to remind you that we have plenty of sand avai= lable for educational use (see partial list below). Fall is a great time to volunteer to help teachers. Microscopic Explorations is a great hands = on science program that teaches microscopy and shows what a useful tool microscopes are for many other fields of science. Students, teachers, parents, and volunteers all enjoy and learn from this program. If you = would like to learn more about Project Micro and Microscopic Explorations ch= eck out the Project Micro web page at: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html. If you wou= ld like to request sand for educational use, contact me at:
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com
SAND LOCATION Golden sand Patuxent River Chesapeak Bay, MD Beige dune sand Lake Michigan, Indiana Dunes, IN Beige dune sand Lake Michigan, Sleeping Bear Dunes, Empire, MI Red rock sand San Francisco Bay, CA Beige ocean sand Monterey Beach, CA Gray ocean sand T Street Beach, San Clemente, CA Beige pebble sand Waikiki, HI Beige shell sand Waimea Bay, HI Green beach sand Big Island, HI White ocean sand Cancun, Mexico Pink coral sand Barbados White ocean sand Marthon, Florida (gulf side) Orange beach sand Saudi Arabia Beige ocean sand Valparaiso, Chile Black ocean sand Curico, Chile Brown ocean sand Castelldefels, Spain Golden ocean sand Sydney, Australia Golden desert sand Cairo, Egypt Beige ocean sand Adelaide, Austrailia Beige ocean sand Nassau, Bahamas Black mineral sand Antartica =
A short time ago I inquired after recommendations for full-page, color, printers, including Sony, Kodak, Fuji, and Codonics machines. Today I had a demo of these printers, plus the Tektronix Phaser 840. I was intrigued by the 840s image quality and cost for purchase and operate.
Can anyone comment on this printer? Perhaps off-list would be best.
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"Cargille Optical Liquids and Mounting Media for the Microscopist "
Speaker is Robert Sacher of Cargille Laboratories.
Robert Sacher will speak about Cargille Refractive Index Liquids and the different techniques for using them. He will discuss the Cargille Microscope Immersion Oils and there fluorescence and viscosities. Plus he will discuss the Cargille Meltmount Mounting Media and their different refractive indices. Issue of toxicity will be considered as well as the significance of the Becke line, etc. This is a great opportunity to learn basic information about optical liquids and mounting media; understanding how to use these properly is a key to obtaining the best possible image through a light microscope. There will be ample time for questions and answers
We are considering replacing one of our old EDS systems (interfaced to a TEM) with a new one. I would appreciate comments/opinions from USERS of any of the following systems .(Please respond privately to : Jordi.Marti-at-AlliedSignal.com):
PGT ....IMIX EVEX ...VIDX EDAX...Falcon
We are primarily interested in the following:
UTW, Si-detector. 30sq.mm. Quant (Thin film). Mapping Windows NT based system.
I'm looking for the spec. or sample source code (c, c++) for reading and writing the EMSA x-ray spectra file format. I've looked on the msa ftp site but could not find a reference to it. Since this is for a new SEM/EDX application that we are writing, I would also be interested in including other x-ray spectra file formats that users might require. If you have a request (and the format is public), then let me know.
Thanks Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
} } } I'm looking for the spec. or sample source code (c, c++) for } reading and writing the EMSA x-ray spectra file format. I've looked on the } msa ftp site but could not find a reference to it. ............................................................................... .
Abstract Deadline for papers and posters is next Friday October 15th. Please follow exactly the abstract instructions in our circular or website.
[Hint for those who hate laying out formatted abstracts - look at the conference website (www.anu.edu.au/EMU/acem) and click the Abstract Instructions button. There you can download an abstract document already laid out in MS Word. Should save you heaps of time!] **************************************************************************** ----------------------------------------------- Dr Marion A. Stevens Kalceff Australian Research Fellow (ARC), MAU, Faculty of Science, University of Technology, Sydney PO Box 123 Broadway NSW 2007 Australia Tel +61 2 9514 1702 (lab) +61 2 9514 1621 (office) Fax b+61 2 9514 1703 email: marion-at-phys.uts.edu.au Marion.Stevens-Kalceff-at-uts.edu.au -----------------------------------------------
The most commonly used procedure for pre-embedding immunogold labeling is: 1. fixation, 2. permeabilization, 3. blocking, 4. primary antibody incubations, 5. secondary antibody incubation (ultrasmall gold particles conjugated probes), 6. glutaraldehyde post-fixation, 7. silver enhancement, 8. EM processing and embedding. The quality of ultrastructure and the degree of antibody penetration are mainly the result of the choice of fixatives and permeablization reagents. The type of fixative is very much limited by the vulnerability of the epitope configuration to the fixative. Unfortunately, there is no set formula for determining the optimal fixative for each protein. It needs to be tested empirically. If the above were the only concern, one would use the strongest fixative consistent with maintenance of epitope configuration. However, when the epitope is "tough", one still should consider the effect of fixation on antibody penetration. Stronger fixation causes tighter cross-linking of cellular components, which hinders antibody penetration. In reality, the effects of fixation on antigenicity and antibody penetration are often dealt with as a whole. The common practice is to try several different fixatives with the permeabilization method of choice. In my experience, 4% paraformaldehyde plus 0.2% glutaraldehyde is a good place to start for many receptor and transporter proteins in neuronal samples. The common permeabilization methods are saponin and Triton X100. (some also use the freeze-thaw method and the "dehydrate-rehydrate" method). I have heard people say they got better ultrastructure in cell culture with saponin than with Triton, but I do not know if anyone has done a comparison of ultrastructure quality versus labeling intensity in a quantitative way. It has been suggested that the membrane-permeabilizing effect of saponin is due mainly to its property of forming complexes with membrane-associated cholesterol (M. Wassler, 1987, Schlosser & Wulff, 1969). The application of saponin as a permeabilizing reagent in pre-embedding immunogold labeling should be continuous throughout all immunoreagent incubations and washing steps between incubations. The concentration usually is around 0.05%. Triton is a non-ionic detergent that disrupts hydrophobic associations. When applied to membranes, it not only destroys lipid bilayers, it can also solubilize membrane bound proteins, especially if used in high concentration. For the latter reason, the use of Triton should be gentle, or even avoided if necessary, when using pre-embedding immunogold labeling for receptor and transporter proteins. For 50 um brain vibratome sections, I usually use 0.05% Triton before the blocking step for 30 min; for cell culture, 5-10 min. It should also be mentioned that 0.1% sodium borohydride that is often used in pre-embedding immunolabeling for reducing residual aldehyde may also help to "loosen up" the sample for antibody penetration due to its bubbling effect (undocumented personal experience ;-) ). I have used the fixative mentioned above and Triton as the permeablization reagent in both cell culture and brain sections with good preservation of polyribosomes and immunogold labeling of Fragile X mental retardation protein, a mRNA binding protein. There are mainly two categories of gold conjugates: colloidal gold conjugates and so called "gold compound" conjugates (Nanogold). I have obtained comparable results with both in many of my experiments. There are several recipes for making silver enhancement solution (Danscher,1981, Burry, 1992) and several silver enhancement kits commercially available. But in terms of enhancement efficiency, ultrastructral "friendliness", and practicality, the kit by Aurion, SE-EM is at the top on my list for EM level pre-embedding immunogold labeling. I don't think one is allowed to send attachments to the server. But if you like, I can send two protocols (one for cell culture, the other for brain sections) to you at your own email address so that you have a place to start.
I don't mean to write a book here. Hope it helps.
Hong ================ Hong Yi Emory University School of Medicine Neurology Microscopy Core Laboratory Rm 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
I tried to do this a few years ago and I would like to warn you about an important factor. A friend of mine called me up and said they had made an excursion and found a small crawfish. He asked me to do something with it. Actually, = it wasn't that small (about 20 cm). In spite of this I made a box out of = plexiglass, and tried to embed it into Araldite which was used in our lab to embed our metallic samples to get polished cross sections. I fixed the legs of the crawfish with a little glue in the box, then I = made the Araldite mix and poured it over the crawfish immediately. I put the = whole thing on a desk over some typing paper and left it in the room where our microscope was situated. I left it alone only for a few minutes. When I returned back to see what had happened I realized that the Araldite had already solidified but in a manner I did not like. It was = so hot that the lacquer layer under it had been burnt. The paper could have caught fire! The Araldite itself had become opaque and yellow. So if you mix Araldite in larger quantities, keep stirring and cooling it for a while before = pouring it over your special stuff.
Laszlo Varga
-----Eredeti =FCzenet----- Felad=F3: Jon Krupp [SMTP:jmkrupp-at-cats.ucsc.edu] K=FCldve: 1999. okt=F3ber 7. 22:22 C=EDmzett: Microscopy-at-Sparc5.Microscopy.Com T=E1rgy: Plastic embedding for display
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Hi:
Does anyone know how to make those clear plastic blocks with embedded specimens? Sometimes you see them with flowers or bugs inside used as paperweights. We would like to make some with some special stuff inside = for display in our lab.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Just found an online copy of the Philips info (Now FEI Beam Technology). It's at http://www.feibeamtech.com/schottky/schottky.htm
Regards
Tim
*********************************************************** EuroFE Field Emission Network A Network of the European Science Foundation http://www.esf.org/ Tim E. Harper EuroFE Network Co Chairman CMP Cientifica s.l Tel +34 91 640 71 85 Fax: +34 91 640 71 86 http://www.cmp-cientifica.com/Eurofe
-----Original Message----- } From: Hooghan, Tejpalkaur K (Tejpalkaur) [mailto:hooghan-at-lucent.com] Sent: Friday, October 08, 1999 10:08 PM To: 'tim-at-cmp-cientifica.com'
Hello all
On several occasions there has been discussion=20 about the most economical way to update a broken=20 EDS-system. We have a repaired detector (with Be-window) for=20 our JEM1200EX but the computer unit got to the=20 end of its road. There are no spare parts=20 awailable for the TN2000 anymore. Could someone suggest an economical way to=20 upgrade the system so that we could continue=20 analysing samples.
Thanks in advance, Jouko
Jouko M=E4ki University of Turku Laboratory of=20 Electron Microscopy PhD Kiinamyllynkatu 10 FIN-20520 TURKU=20 FINLAND Laboratory Manager Tel.: +358 2 333 7318=20 +358 40 505 2521 E-Mail: jouko.maki-at-utu.fi Fax: +358 2 333 7380
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Mary, Check out the reference: Howard, R.J. and K. L. O'Donnell (1987) Freeze Substitution of Fungi for Cytological Analysis. Experimental Mycology 11:250-269.
This is only one of a number of papers by Rick Howard and others describing Freeze-substitution protocols. I have been using a very similar technique for the last 10 years with great success. Please contact me directly if you need additional information.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
} } } Can anyone give me some refernces for a method of fixing tissue in liquid } propane, then dehydration in acetone at -80C ? I think that is the } sequence. Anyway, a friend was telling me about this method and now I } cannot get in touch with him to find out more. Thanks Mary } } John P.B. & Mary } mpfauth-at-teleport.com } } }
Dear Listservers, Does anyone have a reference for some TEM electronmicrographs of trematodes? To be more specific, we are studying intestinal fluke infections within South Australia and are interested in the ultrastructure of the cercarial, metacercarial and adult stages of the trematode family Brachylaimidae which is part of the wider group called "Digenea". Thankyou. John Brealey EM Unit The Queen Elizabeth Hospital Adelaide South Australia
Wow! Get a hold of this software and try it. They have a demo version that does all the functions and times out after a couple of months. This is pretty awesome software. From what I can see, it will pull out detail from underexposed negs. To the eye, the negs look underexposed but to the Lucis software there is information there. The better the scanning function to digitize the neg the better the results will be.
I'm working with a demo version of Lucis right now and I am becoming more and more impressed each day.
gary g.
} X-Sender: mme-at-mail.map.com } X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32) } Date: Wed, 06 Oct 1999 11:49:18 -0400 } To: Donald Delaney {delaneyd-at-mcw.edu} , microscopy-at-sparc5.microscopy.com } From: Barbara Foster {mme-at-map.com} } Subject: Re: Photoshop Negative Processing } } } Don, } } There is some new software, called "Lucis" which is really great for this } purpose. Suggest that you give them a shout at Image Content Technologies, } 860-223-4710 or wwww.imagecontent.com. Ron Lund is VP sales. } } Hope this is helpful. } } Best regards, } Barbara Foster } Consortium President } Microscopy/Microscopy Education ...Educating microscopists for greater } productivity. } } } } } } I have some negatives taken on the TEM which developed extremely light. I } } was wondering if anyone knows any tricks on photoshop for generating good } } quality prints. I typically adjust the levels and condense the size of the } } picture, but I am still not completely happy about the quality of the } prints. } } } } Don Delaney } }
Our laboratory has recently sold our old TEM and EDX unit (circa 1976). The new owner did not want all the electronic parts, so rather than throw them in the dumpster, I am offering them to anyone who can use them. All I ask is you pay the shipping costs.
The parts leftover are:
Control console model 5372 for a Kevex 5500 EDX Hitachi monochrome video display monitor for above Polaroid camera for the display monitor
Polaroid camera unit for JEOL 100C TEM Electronic panel (for control of parameters like focus, magnif, etc) and all associated boards for JEOL 100C TEM
If you have any questions, please contact me off-list.
Helen Mayer UCAR Carbon Company Parma, OH 216-676-2373
In regards to the thread on Lucis software and its improvement of underexposed negs (see below), I offer this message I got along with the demo of the software (I haven't had time to try it):
Dear Tom, My name is Ron Lunn. I manage sales in microsopcy and analytical imaging. Lucis is available as a stand alone program for PCs. Currently we are encouraging users to try Lucis by offering a $1,000 discount if a purchase order is received at ICT by 11/30/99. So the price of Lucis would be $1,495. After 11/30/99, the price reverts to $2,495. I have attached demonstration software that you may use for 2 months. It will time out. The User's Guide is available from our web site, imagecontent.com. Lucis is very easy to use.
Thank you for your interest. Please let me know if I can be of further assistance. I am using our T1 line to send you the software (the bwilliams-at-imagecontent.com address). I can be reached at 860-435-0194 or lucis-at-mohawk.net. I will follow up with a phone call to discuss your specific application. Ron Lunn Sales, Microscopy & Analytical Imaging
Image Content Technology LLC 185 Main Street, Suite 211 New Britain, CT 06051 tel: 860-223-4710 fax: 860-229-8164 bwilliams-at-imagecontent.com
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Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I was just recently told that Kodak made a change in the dye used in their blank CDs for making CD-ROMs, and that this change is causing compatibility problems with some CD burners.
Has anyone experienced this problem? What brands of blank CDs are folks using these days for burning archival CDs? Note, I'm referring to CD-ROMs only *not* CD-RWs and the like. Nor DVDs.
Thanks!
Phil
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 USA Address for courier deliveries: 6319 Pheasant Lane #A-12 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net
Dear Renata, I completed my Ph.D. project working on cercarial sensory receptors. One of the techniques I employed was TEM and I did an extensive work with cer- cariae of families Allocreadiidae, Lecithodendriidae, and Opecoelidae. As a pilot project, I did some SEM work with sporocysts of Leucochloridium which is closely related to your digeneans. TEM is a tricky business, involving a lot of perseverance and patience. A good reference is Pojmanska and Machaj, 1991. Differentiation of the ultrastructure of the body wall of the sporocyst of Leucochloridium paradoxum. International Journal for Parasitology 21 (6): 651-659. I got better results working with acrolein, a fast penetration che- mical that improves fixation of nervous tissue but very dangerous (it would be wise to avoid unnecessary risks). There are other means to improve fixation such as to cut the specimens into large pieces or fix them under coverslip ( penetration is far improved in flat specimens). These procedures are size- dependent though, but I encourage you to try both of them. Hope this info helps you. Good luck with your project and don't hesitate to contact me in case I could be of further assistance.
Check your library to see if it has or can get the series "Microscopic Anatomy of Invertebrates" Fred Harrison series editor, published by John Wiley-Liss in the US. 15 volumes. I forget which one the trematodes is in, but it's one of the earlier volumes.
Phil
} Dear Listservers, Does anyone have a reference for some TEM } electronmicrographs of trematodes? To be more specific, we are studying } intestinal fluke infections within South Australia and are interested in } the ultrastructure of the cercarial, metacercarial and adult stages of the } trematode family Brachylaimidae which is part of the wider group called } "Digenea". Thankyou. John Brealey EM Unit The Queen Elizabeth Hospital } Adelaide South Australia
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 USA Address for courier deliveries: 6319 Pheasant Lane #A-12 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net
I think you have some problems with your hot cathod instrument. I use mine mostly at 5-10 kV with routine magnifications up to 50k (and somtimes 100-150k). For beam sensitive samples I use high voltage down to 0.4kV. Contrast is excellent. Wet mode and EDS are fine too. So, I believe, unconvinient cold emission instruments should be used only for magnifications above 100k.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Phone: (816) 235-2072 Fax: (816) 235-5524
} -----Original Message----- } From: Bruce E. Brinson [mailto:brinson-at-rice.edu] } Sent: Thursday, October 07, 1999 9:01 PM } To: Stephen McCartney } Cc: Microscopy-at-Sparc5.Microscopy.Com } Subject: Re: cold field emission vs. Schottky emitters } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } HI Stephen, } } We have one of each. Hands down the cold cathode } instrument is the superior } imaging tool but that instrument is optimized for hi-res } imaging. Our hot } cathode instrument is a analytical platform with poor vacuum } / high vacuum } modes. I believe a local space agency bought the next } generation of our cold } cathode SEM after a demo on ours. They were using a hot } cathode instrument } previously. Interestingly we bought a similar hot cathode } instrument later but } only because that is how the system came. It was purchased } for it's poor vacuum } capability. } } I use the cold cathode instrument for all (imaging) but } the lowest mag work } ( {2k). It suffers from spherical aberration at lower mags. } This is my SEM of } choice. } I use the hot cathode instrument for low mag work ( {2k), } EDS or wet work. } It has a larger area of view. At times I have used it up to } x10K but only } because I was there & had a favorable specimen. } } The contrast & image detail of the cold cathode system is } decidedly } superior. I can also point out that the images can be } acquired at 5 kV, some } times less. I always use 30 kV for the hot cathode SEM. } } Let's be clear....There are differences in the optics & } secondary detection } systems which account for some of the difference in image quality. } Both instruments have a place. } } On reliability, the cold cathode went on line in 1994. } The instrument is } anvil reliable. The hot cathode has been changed 2-3 times } since 1995. This may } have been a production yield problem. The current one has } been in a while. } } I have purposely avoided using brand names & model #s } because people tend } to associate properties or opinions of individual instruments } with entire } product lines. If you would like specifics, please contact me } off line. } } Meanwhile if your looking to buy, I suggest that you have } vendors image a } variety of materials before you make a firm deci$sion. Also } conditions of final } payment should include being able to replicate those images } on you instrument. } } Bruce Brinson } Rice U. } } } Stephen McCartney wrote: } } } } -------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } On the tread of FE-SEM emitters I would be very interested in hearing what } people have to say about the differences between cold field emitters and } Schottky emitters. I know what the manufacturers say but what about expert } users? Thanks for any input. } } ------------------------------ } Stephen McCartney } Research Associate } Virginia Tech } Materials Institute } 2108 Hahn Hall } Blacksburg, VA 24061-0344 } USA } } TEL: 540-231-9765 } FAX: 540-231-8517 } ------------------------------
} } From: Rhea Freeman {freemanr-at-bio.indiana.edu} } } Subject: LM,TEM,SEM Job Announcement } } } } } JOB ANNOUNCEMENT } **************** } The Indiana Molecular Biology Institute at Indiana University is seeking a } Ph.D level scientist with demonstrated excellence in microscopy to direct } the Molecular Biology Microscopy Facilities, which include light, confocal } fluorescence, transmission, and scanning electron microscopes. The } position includes overall management of the facilities, user training, and } user supervision. We seek a candidate who will be an active participant } with Institute faculty in the development of our microscopy capabilities } and training programs. In addition, pursuit of independent and } collaborative research will be strongly encouraged. Salary: $37,000. } } Candidates should send a resume and have three letters of reference } sent to Dr. Rudolf A. Raff, Director, Indiana Molecular Biology Institute } Indiana University, Jordan Hall, 1001 E. Third St., Bloomington, } IN 47405. Deadline for application: Nov. 1, 1999. Indiana University is an } Equal Opportunity/Affirmative Action Employer. } } Additional information is available at } http://www.bio.indiana.edu/research/imcb/MFM_Ad.html } } ------------------------------------------------------------------------------- } } Rhea Freeman Ph. 812-855-4183 } Administrative Assistant Fax 812-855-6082 } Indiana Molecular Biology Institute } Jordan Hall 322A } Indiana University } Bloomington, IN 47405 }
Thanks for letting us know that you have not tried this product.
Will you post something worthwhile after you have tried it?
gg
At 02:40 PM 10/11/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } In regards to the thread on Lucis software and its improvement of underexposed negs (see below), I offer this message I got along with the demo of the software (I haven't had time to try it): } } Dear Tom, } My name is Ron Lunn. I manage sales in microsopcy and analytical imaging. } Lucis is available as a stand alone program for PCs. Currently we are } encouraging users to try Lucis by offering a $1,000 discount if a purchase } order is received at ICT by 11/30/99. So the price of Lucis would be } $1,495. After 11/30/99, the price reverts to $2,495. I have attached } demonstration software that you may use for 2 months. It will time out. } The User's Guide is available from our web site, imagecontent.com. Lucis } is very easy to use. } } Thank you for your interest. Please let me know if I can be of further } assistance. I am using our T1 line to send you the software (the } bwilliams-at-imagecontent.com address). I can be reached at 860-435-0194 or } lucis-at-mohawk.net. I will follow up with a phone call to discuss your } specific application. } Ron Lunn } Sales, Microscopy & Analytical Imaging } } } } Image Content Technology LLC } 185 Main Street, Suite 211 } New Britain, CT 06051 } tel: 860-223-4710 } fax: 860-229-8164 } bwilliams-at-imagecontent.com } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Anybody got a JEOL wds spectro or three, to fit 840/733, that they would like a good home for?
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Scanning Electron Microscopist Northwestern University
The electron probe instrumentation center (EPIC) at Northwestern University has an immediate opening for a scanning electron microscopist. EPIC is a part of the world renowned materials research center (MRC) and the department of Materials Science & Engineering at Northwestern.
The scanning electron microscopist would be in charge of all of EPIC SEMs (Hitachi S570, FE SEM S4500 and VP SEM 3500N), their accessories (EDS, EBSD/OIM, LHe stage; systems) and the Hitachi FB-2000A focused ion beam (FIB) system. All microscopes in EPIC are under full service contract. Thus, the duties include mainly training students/users, development of specialized techniques and applications, minor maintenance, record keeping and billing. A BS/MS degree in physical/biological sciences is required. The person must have hands-on experience in all aspects of SEM: specimen preparation, EDS, digital acquisition, processing etc. All levels of experience will be considered. Compensation would commensurates with experience and qualifications.
Send cover letter, resume and three references to:
Prof. Vinayak P. Dravid, Director EPIC Materials Science & Engineering Northwestern University, 3013A MLSB Evanston, IL 60208 E-mail: v-dravid-at-nwu.edu Fax: (847) 491-7820
http://epic.ms.nwu.edu/epic/index.htm
Northwestern University is an Affirmative Action/Equal Opportunity Employer.
Hiring is contingent upon eligibility to work in the United States. *******************************************************
Postdoc Opening in Nanostructured Materials Synthesis and Characterization
A postdoctoral scholar position is immediately available at Northwestern University, Evanston, IL in the area of synthesis and characterization of nanostructured materials.
This project is jointly supervised by Profs. D Lynn Johnson and Vinayak P. Dravid, and concerns with synthesis and characterization of ultrafine elemental, alloy and compound particles via arc evaporation. The aim of the project is to synthesize Au/Al2O3 and related catalytic nanostructure systems as a part of multidisciplinary, multi-departmental activity of the Institute of Environmental Catalysis (IEC).
The position requires a PhD in physical sciences/engineering. Considerable hands-on experience in instrumentation, machine hardware and mechanical design in the context of nanostructure synthesis is required. This may include experience in CVD, PVD or related vapor phase techniques for nanostructured materials. Experience in advanced TEM is highly desirable.
The position is available immediately for at least one year with extension for additional year upon mutual consent. Salary and compensation would commensurate with experience.
HIRING IS CONTINGENT UPON ELIGIBILITY TO WORK IN THE USA.
Please forward resume with three names of references to:
Prof. Vinayak P. Dravid Associate Professor, Materials Science & Engineering 2225 N. Campus Drive, 3013A MLSF Northwestern University, Evanston, IL 60208, USA Fax: (847) 491-7820 E-mail: v-dravid-at-nwu.edu http://nuinfo.nwu.edu/materials/faculty/vpd.html
The correct WWW address for Lucis is: http://www.imagecontent.com/lucis/default.htm
Looking on their pictures I was impressed how Lucis enhanced details in the shadows. They use a special patented algorithm to extract data from the image. So, it is not simply "contrast/brightness" or even "gamma" adjustment. After Lucis we probably will have a deal with new, "treated' image. In some particular applications it may greatly improve the quality of information we can extract from image.
Sincerely,
} Date: Mon, 11 Oct 1999 13:40:34 -0500 } From: Tom Phillips {PhillipsT-at-missouri.edu} } Subject: Re: Underexposed Negatives + Lucis software } X-Sender: PhillipsT-at-pop.email.missouri.edu } To: Microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
We are going for a new analytical electron microscope in Trondheim next year. Anyone who knows about manufacturars of EDX software that includes biological thin film quantificaltion according to the "continuum" (Hall---) method?
Vennlig Hilsen=20 dr.ing Gunnar Kopstad overingeni=F8r Avd f Patologi, Rit
On July 21, I purchased 200 spindle packed, unbranded Kodak gold/gold CD-R blanks. FWIW, One S/N in the series is 9072 3101 2254. Don't know how to "read" any date... FYI, since I bought spindle packs ( {$), and to better fit in (real) file drawers, I found a supplier of tyvek CD envelops. $75 per 1000 windowless envelopes w/flap sure beat the "crystal" price and adds no bulk.
[...Sorry about weird margins. Maybe one day IT will dump this awful email program they make us use]
My CD-R is a Smart & Friendly 6x TurboWriter (SCSI) installed on my IXRF EDS PC (Do you speak acronyms? :).
These CDs (so far) have been read by about 10 different PCs here. I have had no problem in either writing or reading, but these may have been produced prior to the possible change you mentioned.
Will be interested in developments.
Regards, Woody White McDermott Technology, Inc. Lynchburg Research Center ---------------------------------- I was just recently told that Kodak made a change in the dye used in their blank CDs for making CD-ROMs, and that this change is causing compatibility problems with some CD burners.
Has anyone experienced this problem? What brands of blank CDs are folks using these days for burning archival CDs? Note, I'm referring to CD-ROMs only *not* CD-RWs and the like. Nor DVDs.
Thanks!
Phil
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 USA Address for courier deliveries: 6319 Pheasant Lane #A-12 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net
If you had read my posting, you would have seen it contains the new information that the software costs $1495. For some people, the cost of a product is a major factor in whether it is worth demo'ing.
} Huh? } } Thanks for letting us know that you have not tried this product. } } Will you post something worthwhile after you have tried it? } } gg } } At 02:40 PM 10/11/99 , you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } In regards to the thread on Lucis software and its improvement of } underexposed negs (see below), I offer this message I got along with } the demo of the software (I haven't had time to try it): } } } } Dear Tom, } } My name is Ron Lunn. I manage sales in microsopcy and analytical imaging. } } Lucis is available as a stand alone program for PCs. Currently we are } } encouraging users to try Lucis by offering a $1,000 discount if a purchase } } order is received at ICT by 11/30/99. So the price of Lucis would be } } $1,495. After 11/30/99, the price reverts to $2,495. I have attached } } demonstration software that you may use for 2 months. It will time out. } } The User's Guide is available from our web site, imagecontent.com. Lucis } } is very easy to use. } } } } Thank you for your interest. Please let me know if I can be of further } } assistance. I am using our T1 line to send you the software (the } } bwilliams-at-imagecontent.com address). I can be reached at 860-435-0194 or } } lucis-at-mohawk.net. I will follow up with a phone call to discuss your } } specific application. } } Ron Lunn } } Sales, Microscopy & Analytical Imaging } } } } } } } } Image Content Technology LLC } } 185 Main Street, Suite 211 } } New Britain, CT 06051 } } tel: 860-223-4710 } } fax: 860-229-8164 } } bwilliams-at-imagecontent.com } } } } } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Wow! Get a hold of this software and try it. They have a demo version } } } that does all the functions and times out after a couple of } months. This is } } } pretty awesome software. From what I can see, it will pull out detail from } } } underexposed negs. To the eye, the negs look underexposed but to the } } } Lucis software there is information there. The better the } scanning function } } } to digitize the neg the better the results will be. } } } } } } I'm working with a demo version of Lucis right now and I am becoming more } } } and more impressed each day. } } } } } } gary g. } } } } } } } } } } X-Sender: mme-at-mail.map.com } } } } X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32) } } } } Date: Wed, 06 Oct 1999 11:49:18 -0400 } } } } To: Donald Delaney {delaneyd-at-mcw.edu} , microscopy-at-sparc5.microscopy.com } } } } From: Barbara Foster {mme-at-map.com} } } } } Subject: Re: Photoshop Negative Processing } } } } } } } } } } } } Don, } } } } } } } } There is some new software, called "Lucis" which is really great for this } } } } purpose. Suggest that you give them a shout at Image Content } Technologies, } } } } 860-223-4710 or wwww.imagecontent.com. Ron Lund is VP sales. } } } } } } } } Hope this is helpful. } } } } } } } } Best regards, } } } } Barbara Foster } } } } Consortium President } } } } Microscopy/Microscopy Education ...Educating microscopists for greater } } } } productivity. } } } } } } } } } } } } } } } } } } I have some negatives taken on the TEM which developed } extremely light. I } } } } } was wondering if anyone knows any tricks on photoshop for } generating good } } } } } quality prints. I typically adjust the levels and condense } the size of the } } } } } picture, but I am still not completely happy about the quality of the } } } } prints. } } } } } } } } } } Don Delaney } } } } } } } } } Thomas E. Phillips, Ph.D. } } Associate Professor of Biological Sciences } } Director, Molecular Cytology Core Facility } } } } 3 Tucker Hall } } Division of Biological Sciences } } University of Missouri } } Columbia, MO 65211-7400 } } (573)-882-4712 (voice) } } (573)-882-0123 (fax)
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Postdoc Opening in Advanced Analytical TEM and Electron Holography
A postdoctoral scholar position is immediately available at Northwestern University, Evanston, IL in the area of advanced Analytical TEM of nanostructured materials.
This project is being supervised by Prof. Vinayak P. Dravid, and concerns with analysis of nanoscale chemistry, structure and electrostatic fields at interfaces and junctions in advanced microelectronics using analytical TEM and holography techniques.
The position requires a PhD in physical sciences/engineering. Considerable hands-on experience in advanced TEM techniques such as HRTEM, EDS/EELS, CBED and computation/simulations is required. Experience in electron holography is highly desirable.
The position is available immediately for at least one year with extension for additional year upon mutual consent. Salary and compensation would commensurate with experience.
HIRING IS CONTINGENT UPON ELIGIBILITY TO WORK IN THE USA.
Please forward resume with three names of references to:
Prof. Vinayak P. Dravid Associate Professor, Materials Science & Engineering 2225 N. Campus Drive, 3013A MLSF Northwestern University, Evanston, IL 60208, USA Fax: (847) 491-7820 E-mail: v-dravid-at-nwu.edu http://nuinfo.nwu.edu/materials/faculty/vpd.html *******************************************************
Metropolitan Microscopy Soc. -- Fall Meeting 1999 =================================================
Time: 9:00 am (registration begins) **PLEASE PRE-REGISTER** It is im- portant. (See below)
Place: Zeiss, Thornwood, NY (914) 747-1800.
} From most locations, follow your choice of major highways to I-287 and the Saw Mill River Parkway. Take the Saw Mill north. From the nearby north, follow the Taconic Parkway south and take the Saw Mill Parkway north.
Follow the Saw Mill to Exit 27 (Marble Ave.). Take a right onto Mar- ble Ave. Follow Marble through the underpass and traffic light. Bear right at the top of the hill (becomes Columbus Ave.). Zeiss Drive is on the left, just past the Rose Hill shopping center.
-------------------------------------
AGENDA
9:00 - 9:45 : Registration (Coffee)
9:45 - 10:00 : Introductory Remarks and society business (Al Sicignano).
10:00 - 10:45 : "Imaging Microstructure Using Orientation and Chemis- try, Paul Manwairing, TSL
10:45 - 11:30 : "Automated EBSP and Orientation Imaging for Micro- structural Characterization of Ti and Ni based Superalloys ", John Sutliffe, GE Corporate R& D.
1:45 - 3:15 : "Image Analysis and Image Processing", John Russ, Professor Materials Science and Engineering Dept. NC State University, Raleigh, NC
-------------------------------------
PRE-REGISTRATION INFORMATION
Zeiss Corp. has graciously agreed to host our meeting at their facility in Thornwood, NY. Due to the nature of their corporate fa- cility, IT IS ESSENTIAL THAT MEMBERS PRE-REGISTER so that an attendee list can be delivered to the security people at Zeiss for generation of guest badges.
**THE PRE-REGISTRATION DEADLINE IS Oct. 15th** and can be accom- plished electronically. Please respond via email or fax to Evan Slow directly. A simple email note or fax is all that's required to pre-- register. You can then bring the required fee with you to the meet- ing.. For pre-registrants, the meeting fee, which includes lunch, will be $15.00.
***ON-SITE REGISTRANTS WILL BE CHARGED $25.00***
Pre-register with Evan Slow --- (201) 760-2525 (FAX) ess-at-feico.com
Philip L. Flaitz IBM Analytical Services, Hopewell Junction, NY http://www.chips.ibm.com/services/asg Ph.......(914) 892-3094, FAX -2003 flaitz-at-us.ibm.com
by aretha.jax.org (8.9.1/8.9.1) with SMTP id NAA15628 for {microscopy-at-sparc5.microscopy.com} ; Tue, 12 Oct 1999 13:20:15 -0400 (EDT) Message-Id: {3.0.5.32.19991012132545.008d7180-at-aretha.jax.org} X-Sender: lsb-at-aretha.jax.org X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.5 (32)
Thanks to everyone who gave me their tips for preparing intestines for SEM. The super-dried acetone did the trick. The results will show up in "Science" sometime and the investigator was very pleased! Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Hello, A colleague of mine is trying to disperse membrane proteins for negative staining with different types and amounts of detergents, but so far without success (proteins aggregating). I was thinking of lightly fixing the prep before extracting the lipids, then using detergent to extract all lipids, followed by negative stain. We would appreciate comments on this or any suggesstions are welcomed. Thank you.
Is there anyone who has a mannual for E5400 Polaron sputter coater.I would be greatful to you if you could fax a copy.Also I need to find cheaper place to buy coating target source.
Thank you
Reena Zalpuri EM Lab UC Berkeley =46ax 510-643-6207 E-mail jubu-at-uclink4.berkeley.edu
Philip Oshel had questions regarding the suitability of Kodak Recordable CDs for archiving images. My lab uses the KODAK CD-R GOLD ULTIMA Recordable CD without problesm. Kodak supplies a whole family of products, one of which should give superior performance in most CD recorders. Interested microscopists can find specific information on the Kodak web site at
http://www.kodak.com/US/en/digital/cdr/
Interested microscopists can also get help for choosing the correct media for their recorder from the Kodak Digital Imaging Support Center 9 am - 8 pm Mon-Fri at (800) 235-6325.
Best Regards
John Minter Analytical Technology Division Eastman Kodak Company Rochester, NY 14652-3712 Phone: (716) 722-3407 FAX: (716) 477-3029 email: minter-at-kodak.com
I had a computer crash, wiped out my "new mail", would the person from South Africa who contacted me please contact me again?
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I misread the msg. My response was out of line and off base.
Sorry about that.
gary g.
} X-Sender: gaugler-at-pop.calweb.com } X-Mailer: QUALCOMM Windows Eudora Pro Version 4.2.0.58 } Date: Mon, 11 Oct 1999 19:43:29 -0400 } To: Tom Phillips {PhillipsT-at-missouri.edu} } From: "Dr. Gary Gaugler" {gary-at-gaugler.com} } Subject: Re: Underexposed Negatives + Lucis software } Cc: MSA listserver {Microscopy-at-sparc5.microscopy.com} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } I was just recently told that Kodak made a change in the dye used in their } blank CDs for making CD-ROMs, and that this change is causing compatibility } problems with some CD burners. } } Has anyone experienced this problem? What brands of blank CDs are folks } using these days for burning archival CDs? Note, I'm referring to CD-ROMs } only *not* CD-RWs and the like. Nor DVDs. }
O.k., Yes, I have run into this. The kodak CDR's with the info guard layer (the 8x ceritified CDR's, Kodak CDR KOD-899-1309) had a problem in our Yamaha 400T writer initially (the media was NOT recognized as valid media). After some digging the problem was known and correctable from yamaha. All it took was a FlashBIOS upgrade to the Yamaha CDR (which also fixed some other problems and was very easy to perform). Also the Yamaha BIOS update did not effect our ability to burn any other brands of CDR media (They still work just fine)
We have NOT seen any problems since, nor have we seen any problems reading the disks in a wide range (and ages) of PC and Mac CD-ROM drives.
And still recommend the Kodak CDR's to all my users for archiving image data (we have seen some severe data degration problems after numerous reads on the cheaper CDR's).
Note: This was actually just after the Kodak Infoguard CDR disks came out in March 1999, so I am not sure if you are discribing a newer issue. But I would recommend checking out the specific CDRecorders manufacturers site for problems with media and recording.
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
I've used about 150 kodak gold infogard CD's in the past year and a half. Apart from a series of coasters caousd by a dying CD writer I've had no problems with writing them. A couple of people were not able to read them but I'm almost certain it was due to very old CD readers. I've switched to a new Plextor 8220 CD-RW and have had no problems at all.
Cheers
Glenn
Glenn ============================================================= Glenn Poirier Tel:(514) 398-6774 MicroAnalytical Laboratory Fax:(514) 398-4680 Earth and Planetary Sciences McGill University glennp-at-eps.mcgill.ca 3450 University St. castaing.eps.mcgill.ca Montreal, Qc H3A 2A7 -- Millenium Hand and Shrimp -- ============================================================== -----Original Message----- } From: Philip Oshel [mailto:oshel-at-terracom.net] Sent: October 11, 1999 3:08 PM To: microscopy-at-sparc5.microscopy.com
I was just recently told that Kodak made a change in the dye used in their blank CDs for making CD-ROMs, and that this change is causing compatibility problems with some CD burners.
Has anyone experienced this problem? What brands of blank CDs are folks using these days for burning archival CDs? Note, I'm referring to CD-ROMs only *not* CD-RWs and the like. Nor DVDs.
Thanks!
Phil
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today PO Box 620068 Middleton, WI 53562 USA Address for courier deliveries: 6319 Pheasant Lane #A-12 Voice: (608) 833-2885 Fax: (608) 836-1969 (please make sure my name is on any fax) oshel-at-terracom.net
At 07:29 PM 10/11/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm with Vladimir. My LaB6 system does a great job at 100KX and 3KV using well-prepared specimens. E. coli fill the screen at about 75KX and the folds and surface details are nice.
With higher KV, I get too many BSE artifacts. From what I can tell, on my system, the E2 point is right about at 3-4KV.
Does anyone know where I can get disk 10 of Harvard Graphics 2.0? Somehow my disk 10 was lost and I have a new computer and want to install my Harvard Graphics 2.0 (for which I have a legitimate license). The trouble is that SPC won't send me one as it is an old program and they are no longer supporting it. I still have it on my laptop, but when I try to copy it onto my desktop computer (using Uninstaller) it will not open. I have a lot of good materials saved in the HG format and cannot open them without installing all the disks. Also, I still think it's a great program. Thanks for your help, HG user.
The Catholic University of America seeks a technician to perform technical duties in support of the Microstructural Characterization Division of the Vitreous State Laboratory. Technician will be a member of a team responsible for the following: heat treat glasses to induce microstructural changes; prepare glasses for SEM observation using standard metallographic technique; analyze samples for phase content using optical, SEM-EDS, and XRD techniques; instruct students in the operation of microscopes and other equipment; monitor condition of instruments; oversee upkeep of the lab; maintain a safe environment; and other assigned duties.
Applicant should hold a minimum of a bachelor's degree in materials science or related physical science and/or 4+ years related work experience. Candidates should be familiar with electron and optical microscopes, mechanical and electronic equipment, vacuum systems, PC and EDS/WDS systems. Good communication and interpersonal skills are essential.
Catholic University offers excellent benefits and a vacation package.
Interested candidates should forward resume to:
Andrew C. Buechele, Ph.D. Director, Microstructural Characterization Division Vitreous State Laboratory The Catholic University of America 400 Hannan Hall Cardinal Station Washington, D.C. 20064 Telephone: 202-319-4995 Fax: 202-319-4469
e-mail: andrewb-at-rsrch.vsl.cua.edu
Sincerely yours, Andy Buechele
Andrew C. Buechele, Ph.D. Research Professor Vitreous State Laboratory The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
Email: jekstrom-at-mediaone.net Name: James Ekstrom School: Phillips Exeter Academy
Question: I have an microscope objective lens that reads MEOPTA or MEOPLA, it is hard to distinguish. Do you know anything about the manufacturer/location of this objective. Thank you - Jim Ekstrom
} } } } } 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 INTERNET:jekstrom-at-mediaone.net } Email: jekstrom-at-mediaone.net Name: James Ekstrom School: Phillips Exeter Academy
Question: I have an microscope objective lens that reads MEOPTA or MEOPLA= , it is hard to distinguish. Do you know anything about the manufacturer/location of this objective. Thank you - Jim Ekstrom
I've had the following problem with a Philips XL30 and an attached Oxford ISIS 200 series EDS system since I had them installed two years ago. When I collect digital BSE images through the ISIS system using either digital x-ray mapping mapping (SPEEDMAP) or digital image acquisition (AUTOBEAM) there are horizontal broad dark bands on the image. (I have the Philips BSE detector). SE images are ok, and both SE and BSE images taken directly from the microscope are ok. Philips told me the problem was due to the automatic brightness control on the XL30 not being de- activated when the microscope is externally controlled, and that the problem would be fixed in the next version of their software and that the ISIS software would also need a fix. When I checked with Oxford they have not heard of this! I have both pieces of software on the XL30 pc but have tried the ISIS software on a separate pc but the problem still persists. I have also checked for noise in the mains supply which seems ok, the microscope is on its own circuit and there is no heavy electrical equipment in the building. I also tried several different earth connections for the microscope, and different powerpoints for the ISIS box. I know of one other similar system that does not have this problem. Please contact me if you have a similar setup and let me know if you experience this problem or not. I would of though several people would have the problem if it was software related. If I'm the only one seeing this effect I can assume it must be something on my system.
I've posted this message to both the XL users and MSA listservers so I apologise it you get two copies.
Thanks in advance Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
I love HD too. For that times it was a great program. It seems to me that HD2 was working under Win3.x. If so, it is well possible that this is a DOS program and all files are located in one HD directory. If so, the simplest solution is just copying the entire directory. It looks like "illegal way", but it works even for modern programs like Eudora3.06 or Bryce2. It is because many software developers do not like to share "dll"-s and store everything in the program's directory. In your particular case you may choose WinZip (pkzip, may be - old DOS friend) or something like that to create multiple-diskette archive of your HD directory and transfer data to your desktop computer (you will create the same directory from archive on your desktop). When you will try to run the program on the desktop computer, HD, may be, will ask you some files. You should find that files on your laptop ("Find" from Start Menu) and transfer to the same directory on the desktop computer. May be it will take a few "runs".
Good luck.
} Date: Tue, 12 Oct 1999 17:23:41 -0500 } From: Joyce Craig {j-craig-at-CSU.EDU} } Subject: Harvard Graphics Disks } X-Sender: zaluzec-at-microscopy.com } To: Microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
It is unbelievable: such duties for $37K?! There, at UCLA $37K is a level of very moderate post-doc. To be equally perfect in the "light, confocal, fluorescence, transmission, and scanning electron microscopes" just impossible. After 15 years working with EM I would say : I am moderately "perfect" in some very narrow TEM area. This is reason I will not be a successful candidate for that position as well as for many others.
Sergey
} Date: Mon, 11 Oct 1999 18:37:28 -0500 } From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.microscopy.com} } Subject: LM,TEM,SEM Job Announcemen } To: Microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
} Email: jekstrom-at-mediaone.net } Name: James Ekstrom } School: Phillips Exeter Academy } } Question: I have an microscope objective lens that reads MEOPTA or MEOPLA, } it is hard to distinguish. } Do you know anything about the manufacturer/location of this objective. } { http://www.meopta.cz/products/ is a good bet Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK 405 624-2855 GMT -6:00 www.couger.com/gcouger
It's possible to vary the hardness of epoxies by varying the composition. I would have thought that any low viscosity system would have been fine. When I did this, I used Lowicryl - gave good TEM images.
Alun.
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
Reena Zalpuri wrote: =================================================== Is there anyone who has a mannual for E5400 Polaron sputter coater.I would be greatful to you if you could fax a copy.Also I need to find cheaper place to buy coating target source. =================================================== The worldwide central authority and source for these manuals for the older units of the Polaron line is the following:
VG Microtech The Birches Industrial Estate Imberhorne Lane East Grinstead West Sussex RH19 1UB UK Tel: #44(0)1342 327211 Fax: #44(0)1342 315074 E-mail: mwombwell-at-vgmicrotech.com Attn: Mike Wombwell, Manager
Cheaper coater targets? I don't know to what degree they are "cheaper" but at least some of the suppliers of electron microscope supplies and accessories, such as SPI Supplies, offer a full range of replacement sputter coater cathodes. There has been some past discussion on the importance of high purity. We don't know how important that is for you, however, we supply the gold as 0.99999 purity. There is also a variation in thickness in the cathodes from different sources, so in some instances a cathode is more expensive (in comparision) in absolute terms becaue one is comparing a 10 mil thickness with a 5 or even 3 mil thickness. We believe it is a better deal for the customer to have a cathode that is thicker rather than thinner because the "fabrication costs" of a thick one are not different from that of a thin one. You can reduce your costs further by taking advantage of a spent cathode recycling program and in the process, earn a discount on your new cathode purchase. The SPI recycling program is explained on our website, the address given below.
Disclaimer: SPI Supplies is a supplier of replacement cathodes for most brands of sputter coaters used in SEM laboratories and offered the first cathode recycling program in our industry.
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.spi.cc ######################## ============================================
Reena Zalpuri wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Hi, } } Is there anyone who has a mannual for E5400 Polaron sputter coater.I would } be greatful to you if you could fax a copy.Also I need to find cheaper } place to buy coating target source. } } Thank you } } Reena Zalpuri } EM Lab } UC Berkeley } Fax 510-643-6207 } E-mail jubu-at-uclink4.berkeley.edu
Dear Reena,
Sorry we can't help you with the manual, but we do make and sell targets for sputter coaters, including the old Polarons. If you contact me direct with the type of target you want I would be happy to quote.
Thanks,
Deb Sicard -- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
I would like to find a freeware for x-ray spectrum deconvolution, the main problem being the dependance of the apparatus function of a particular Si/Li detector with the energy. Do anybody knows a such freeware. Waiting for your answer, Sincerely, Jean-Michel
Dear Fellow Microscopists, Does anyone located in the central or northern New Jersey area have a sputter coater I can use to coat some tissue culture samples I am preparing for SEM viewing? Our Polaron system is down completely. Please respond off line.
Thank you all in advance,
Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Hoechst Marion Roussel, Inc. 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-hmrag.com
I agree, this is a low salary even here in Dallas, where the cost of living is much lower than in Cal. I hope the employer in Indiana has someone specifically in line for the job or that the cost of living in Indiana is still lower than Dallas. I guess if your an NIH post-doc making 22-24K a year, this could look promising. However, post-docs haven't usually demonstrated excellence in so many areas of microscopy.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } It is unbelievable: such duties for $37K?! There, at UCLA $37K is a level } of very moderate post-doc. To be equally perfect in the "light, confocal, } fluorescence, transmission, and scanning electron microscopes" just } impossible. After 15 years working with EM I would say : I am moderately } "perfect" in some very narrow TEM area. This is reason I will not be a } successful candidate for that position as well as for many others. } } Sergey } } } Date: Mon, 11 Oct 1999 18:37:28 -0500 } } From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.microscopy.com} } } Subject: LM,TEM,SEM Job Announcemen } } 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 } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Direct all inquiries to the address below. } } } } Nestor } } } } ------------------------------------------------ } } } } } } } } From: Rhea Freeman {freemanr-at-bio.indiana.edu} } } } } Subject: LM,TEM,SEM Job Announcement } } } } } } } } } } } } } } } JOB ANNOUNCEMENT } } } **************** } } } The Indiana Molecular Biology Institute at Indiana University is seeking a } } } Ph.D level scientist with demonstrated excellence in microscopy to direct } } } the Molecular Biology Microscopy Facilities, which include light, confocal } } } fluorescence, transmission, and scanning electron microscopes. The } } } position includes overall management of the facilities, user training, and } } } user supervision. We seek a candidate who will be an active participant } } } with Institute faculty in the development of our microscopy capabilities } } } and training programs. In addition, pursuit of independent and } } } collaborative research will be strongly encouraged. Salary: $37,000. } } } } } } Candidates should send a resume and have three letters of reference } } } sent to Dr. Rudolf A. Raff, Director, Indiana Molecular Biology Institute } } } Indiana University, Jordan Hall, 1001 E. Third St., Bloomington, } } } IN 47405. Deadline for application: Nov. 1, 1999. Indiana University } is an } } } Equal Opportunity/Affirmative Action Employer. } } } } } } Additional information is available at } } } http://www.bio.indiana.edu/research/imcb/MFM_Ad.html } } } } } } -------------------------------------------------------------------------- } ----- } } } } } } Rhea Freeman Ph. 812-855-4183 } } } Administrative Assistant Fax 812-855-6082 } } } Indiana Molecular Biology Institute } } } Jordan Hall 322A } } } Indiana University } } } Bloomington, IN 47405 } } } } } } } } } } } } _________________________________ } } Sergey Ryazantsev Ph. D. } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } } } }
Hi everyone, I am looking for a good brand of 1 micron paste to grind TEM sample using Gatan Precision Dimpler. I have used such paste from Buehler, but there is no contact information at hand. Buehler doesn't have a web page at www.buehler.com. Any information in this regard will be appreciated.
Don't forget the cost of living difference between UCLA and Indiana University. Los Angeles is among the top 3 most expensive areas to live in this country, (it is more expensive than Boston by 2-5%). Salaries are generally adjusted to support a standard of living within a community. There exist many many companies that monitor and document Cost of Living in the US and around the world so national and global companies can adjust salaries for employees that get moved around the states and the world.
-geoff
PS It does seem kinda low . . . but then I have heard of tenure track faculty positions that start in the low $40K/year.
Sergey Ryazantsev wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } It is unbelievable: such duties for $37K?! There, at UCLA $37K is a level } of very moderate post-doc. To be equally perfect in the "light, confocal, } fluorescence, transmission, and scanning electron microscopes" just } impossible. After 15 years working with EM I would say : I am moderately } "perfect" in some very narrow TEM area. This is reason I will not be a } successful candidate for that position as well as for many others. } } Sergey } } } } } } JOB ANNOUNCEMENT } } } **************** } } } The Indiana Molecular Biology Institute at Indiana University is seeking a } } } Ph.D level scientist with demonstrated excellence in microscopy to direct } } } the Molecular Biology Microscopy Facilities, which include light, confocal } } } fluorescence, transmission, and scanning electron microscopes. The } } } position includes overall management of the facilities, user training, and } } } user supervision. We seek a candidate who will be an active participant } } } with Institute faculty in the development of our microscopy capabilities } } } and training programs. In addition, pursuit of independent and } } } collaborative research will be strongly encouraged. Salary: $37,000. } } }
} --
Geoff Williams
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
We offer some very high quality (yet reasonably priced) polycrystalline diamond paste that is ideal for use in dimpling. We offer the paste in 1=
micron as well as many other micron sizes. We also offer MicroDi Diamond=
Suspension which is made with the same polycrytalline diamond and is ofte= n used for dimpling and other EM applications. To order, you can call 800-728-2233.
} } } } } 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 Wentao Qin } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hi everyone, I am looking for a good brand of 1 micron paste to grind TEM sample using Gatan Precision Dimpler. I have used such paste from Buehler= , but there is no contact information at hand. Buehler doesn't have a web page at www.buehler.com. Any information in this regard will be appreciated.
This has been an interesting thread. However, I have one gripe. It is not a particular software product which is giving good results with underexposed negatives, but an algorithm which this software uses. It would be nice to bring the discussion to this level.
The problem of making information from a negative visible under challenging circumstances boils down to a rescaling of the pixel values by some function, plus truncations. In addition, filters may be used, which have a more complex dependence than simply the initial value of a given pixel, for instance using its neighboring pixel values. I suspect the good results with Lucis rely on a combination of rescaling and filtering. But does anyone know what Lucis actually does? If not, one might argue that that is bad science: using a black box without knowing what it is doing very often comes back to haunt you. If yes, then it can in principle be done using any software which allows sufficiently low-level processing. I'm not knocking Lucis, but this is supposed to be a scientific forum.
Wharton
+++++++++++++++++++++++++++++++++++++++++++++ Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
} } } } } } } } Wow! Get a hold of this software and try it. They have a demo version } } } } that does all the functions and times out after a couple of } } months. This is } } } } pretty awesome software. From what I can see, it will pull out detail from } } } } underexposed negs. To the eye, the negs look underexposed but to the } } } } Lucis software there is information there. The better the } } scanning function } } } } to digitize the neg the better the results will be. } } } } } } } } I'm working with a demo version of Lucis right now and I am becoming more } } } } and more impressed each day. } } } } } } } } gary g. } } } } } } } } } } } } } X-Sender: mme-at-mail.map.com } } } } } X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32) } } } } } Date: Wed, 06 Oct 1999 11:49:18 -0400 } } } } } To: Donald Delaney {delaneyd-at-mcw.edu} , microscopy-at-sparc5.microscopy.com } } } } } From: Barbara Foster {mme-at-map.com} } } } } } Subject: Re: Photoshop Negative Processing } } } } } } } } } } } } } } } Don, } } } } } } } } } } There is some new software, called "Lucis" which is really great for this } } } } } purpose. Suggest that you give them a shout at Image Content } } Technologies, } } } } } 860-223-4710 or wwww.imagecontent.com. Ron Lund is VP sales. } } } } } } } } } } Hope this is helpful. } } } } } } } } } } Best regards, } } } } } Barbara Foster } } } } } Consortium President } } } } } Microscopy/Microscopy Education ...Educating microscopists for greater } } } } } productivity. } } } } } } } } } } } } } } } } } } } } } } I have some negatives taken on the TEM which developed } } extremely light. I } } } } } } was wondering if anyone knows any tricks on photoshop for } } generating good } } } } } } quality prints. I typically adjust the levels and condense } } the size of the } } } } } } picture, but I am still not completely happy about the quality of the } } } } } prints. } } } } } } } } } } } } Don Delaney } } } } } } } } } } } } Thomas E. Phillips, Ph.D. } } } Associate Professor of Biological Sciences } } } Director, Molecular Cytology Core Facility } } } } } } 3 Tucker Hall } } } Division of Biological Sciences } } } University of Missouri } } } Columbia, MO 65211-7400 } } } (573)-882-4712 (voice) } } } (573)-882-0123 (fax) } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
} I am looking for a good brand of 1 micron paste to grind TEM } sample using Gatan Precision Dimpler. I have used such paste from Buehler, } but there is no contact information at hand. Buehler doesn't have a web } page at www.buehler.com. Any information in this regard will be } appreciated.
Dear Reena, I cannot help you with the manual, but a cheaper source for the target source is: Refining Systems Inc. Abe Dayani PO Box 72466 Las Vegas, NV phone: 702-368-0579 fax: 702-368-0933 You must supply the exact specifications and size, then he will sell you the target at close to the precious metal price.
At 12:44 PM 10/12/99 -0700, you wrote:
} Hi, } } Is there anyone who has a mannual for E5400 Polaron sputter coater.I would } be greatful to you if you could fax a copy.Also I need to find cheaper } place to buy coating target source. } } Thank you } } Reena Zalpuri } EM Lab } UC Berkeley } Fax 510-643-6207 } E-mail jubu-at-uclink4.berkeley.edu
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
The biogeochemistry lab here at Portland State University is in need of a Trimming holder for a Sorval MT-2 Ultra-Microtome. If anyone has a spare holder I would appreciate hearing from you. Direct email to stackc-at-pdx.edu.
Carol Stack Research Assistant -- Department of Geology Portland State University stackc-at-pdx.edu 503.524.8651
I am currently researching new (over the last 10 years) techniques for analyzing fibers with Scanning Electron Microscopy and am having difficulty locating current research/techniques in this area. Any information you could pass on would be extremely helpful. Thanks in advance for your time. Sincerely, David McGregor drmcgreg-at-eos.ncsu.edu
For comparisons on cost of living between locations in the US, check out... http://www.datamasters.com/cgi-bin/col.pl
They report that a person earning 100K in Los Angeles (I know the parties aren't making that much, it's just easier to do percents) will need to earn 85K in South Bend, Indiana to enjoy the same standard of living. Not as big a difference as I thought.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
I like to use a polycrystalline diamond paste instead of the monocrystalline diamond paste. It is more aggressive intially and then breaks down to a smoother size during use. I also prefer the water based stuff instead of the oil based. For dimpling, I use the paste and a one micron suspension slurry as the carrier. They also have smaller size suspension. It works well and it all washes off with water easily.
Wendt Dunnington is my supplier. Phone number is (610) 495-2850 and FAX is (610) 495-2865. You can get a catalog from them.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
---------- } From: Wentao Qin To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi everyone, I am looking for a good brand of 1 micron paste to grind TEM sample using Gatan Precision Dimpler. I have used such paste from Buehler, but there is no contact information at hand. Buehler doesn't have a web page at www.buehler.com. Any information in this regard will be appreciated.
We are just getting started on a project to create a CD ROM with multimedia material to support teaching of SEM, TEM and XRD both for undergrad and graduate students. I would appreciate any advice on publicly available simulation software that could be integrated into the material to provide some interactive bits and pieces.
Does anyone out there have an Evex Analytical VIDX EDS system or a PGT Avalon 8000 EDS system? Could you tell me what you think about the equipment, costumer support, software etc? Is the system user friendly?
If you'd like to tell me directly email me at rgriffin-at-eng.uab.edu
This salary issue is what is sending me out of this field. It is incredulous to think that after 13 years experience in a somewhat high-tech field (microscopy, TEM, SEM, confocal, etc.) it is more lucrative to make ice at the D.U. ice arena, or espresso at Starbucks, than it is to do research in the field of neurobiology. I certainly hope that one day there will be salaries available to allow microscopists to feed their families, pay their mortgage, and live comfortably amongst their neighbors. -Mike Rock my (I wish I had) 2 cents worth
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I first used Kodak CDs about 1 1/2 years ago but switched to the Mitsui brand because at that time Mitsui was making Kodak's gold disks (Mitsui invented the gold formula). There were problems with some CD-R drives due to different disc refectivity and laser beam intensity but as was indicated by Richard E. Edelmann a flashBIOS upgrade usually fixes this problem. I have made hundreds of successful burns with my Yamaha 426 (Smart and Friendly was also using Yamaha drives) and Plextor 412c drives running the most recent flashBIOS upgrades writing to Mitsui gold or silver on gold discs. TDK is another reliable brand and Taiyo discs have a very good reputation but are only available in bulk on spindles. Whatever disks you use keep them cool and dark! A recent pro audio magazine article recommended wrapping the CD-R jewel box with aluminum foil for safest storage.
Phil Oshel wrote: } } } I was just recently told that Kodak made a change in the dye used in their } blank CDs for making CD-ROMs, and that this change is causing compatibility } problems with some CD burners. } } Has anyone experienced this problem? What brands of blank CDs are folks } using these days for burning archival CDs? Note, I'm referring to CD-ROMs } only *not* CD-RWs and the like. Nor DVDs. } } Thanks! } } Phil
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
I ordered my Kodak Infoguard CD-Rs from Computability (.com) at a cost of -at-261 / 200 disks (4 spindle case).
I am trying to locate my order data for the Tyvek envelops (opps!). I do remember that I had to go to the manufacturer even though Staples and Office Max - like stores were listed as distributors. When I called them, I could envision the blank stare from the verbal response... Obtained the location of the mfgr from DuPont Tyvek product support. Will pass along more details if/when I find my PO copy.
Where did you purchase the spindle packed Kodak CDR blanks and how much did you pay?
Also, the tyvek envelopes would be of intrest to me.
Thanks for posting this information. It's just what I have been looking for.
John
} On July 21, I purchased 200 spindle packed, unbranded Kodak gold/gold CD-R } blanks. FWIW, One S/N in the series is 9072 3101 2254. Don't know how to } "read" any date... FYI, since I bought spindle packs ( {$), and to better } fit in } (real) file drawers, I found a supplier of tyvek CD envelops. $75 per 1000 } windowless envelopes w/flap sure beat the "crystal" price and adds no bulk.
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
by mailserv.waikato.ac.nz (8.9.3/8.9.0) with SMTP id KAA92380 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 14 Oct 1999 10:22:30 +1300 Message-Id: {3.0.6.32.19991014101436.0091c510-at-mailserv.waikato.ac.nz} X-Sender: aharris-at-mailserv.waikato.ac.nz X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32)
Our Kevex Delta Plus X-ray analyser has stopped doing dot-maps and is displaying a distorted image in image capture mode (Advanced Imaging). While I would like to purchase a PC-based platform funding makes that unlikely in the forseeable future. Is there anyone out there who has upgraded and has a functional Kevex Delta Plus analyser looking for a home?
Karin, The high-resolution TEM image simulation software TEMPaS (Transmission Electron Microscope Processing and Simulation) that I wrote for the National Center for Electron Microscopy has now been ported to various platforms (Sun, IBM, SGI) under the name of NCEMSS. X-window implementations are available for download from the NCEM at http://ncem.lbl.gov/frames/software.htm -Mike O'Keefe
Karin Pruessner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are just getting started on a project to create a CD ROM with multimedia } material to support teaching of SEM, TEM and XRD both for undergrad and } graduate students. I would appreciate any advice on publicly available } simulation software that could be integrated into the material to provide } some interactive bits and pieces. } } ***************************************************************************** } } Karin Pruessner } Institut fuer Werkstofftechnik } University of Siegen } Paul-Bonatz-Str. 9-11 } D-57068 Siegen } Germany } } Ph: ++49 271 740-2184 } Fax: ++49 271 740-2545 } e: pruessner-at-ifwt.maschinenbau.uni-siegen.de
I agree - and I've just got the demo, looks very interesting. At first glance, particularly for colour, which was an unexpected bonus. Does anybody know what the relationship of the Lucis algorithms are to the ("contrast hysteresis"??) method K-R Peters used a good few years ago..and does anybody have a ref to that website? That method, which at least partly depended on extra hardware processing I think(might well be wrong!) was taken up by JEOL as PIXIE, I think.
This has been an interesting thread. However, I have one gripe. It is not a particular software product which is giving good results with underexposed negatives, but an algorithm which this software uses. It would be nice to bring the discussion to this level.
The problem of making information from a negative visible under challenging circumstances boils down to a rescaling of the pixel values by some function, plus truncations. In addition, filters may be used, which have a more complex dependence than simply the initial value of a given pixel, for instance using its neighboring pixel values. I suspect the good results with Lucis rely on a combination of rescaling and filtering. But does anyone know what Lucis actually does? If not, one might argue that that is bad science: using a black box without knowing what it is doing very often comes back to haunt you. If yes, then it can in principle be done using any software which allows sufficiently low-level processing. I'm not knocking Lucis, but this is supposed to be a scientific forum.
Wharton
+++++++++++++++++++++++++++++++++++++++++++++ Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724 FAX: (208) 533-7863
mailto:wharton.sinkler-at-anlw.anl.gov
} } } } } } } } Wow! Get a hold of this software and try it. They have a demo version } } } } that does all the functions and times out after a couple of } } months. This is } } } } pretty awesome software. From what I can see, it will pull out detail from } } } } underexposed negs. To the eye, the negs look underexposed but to the } } } } Lucis software there is information there. The better the } } scanning function } } } } to digitize the neg the better the results will be. } } } } } } } } I'm working with a demo version of Lucis right now and I am becoming more } } } } and more impressed each day. } } } } } } } } gary g. } } } } } } } } } } } } } X-Sender: mme-at-mail.map.com } } } } } X-Mailer: QUALCOMM Windows Eudora Pro Version 3.0.3 (32) } } } } } Date: Wed, 06 Oct 1999 11:49:18 -0400 } } } } } To: Donald Delaney {delaneyd-at-mcw.edu} , microscopy-at-sparc5.microscopy.com } } } } } From: Barbara Foster {mme-at-map.com} } } } } } Subject: Re: Photoshop Negative Processing } } } } } } } } } } } } } } } Don, } } } } } } } } } } There is some new software, called "Lucis" which is really great for this } } } } } purpose. Suggest that you give them a shout at Image Content } } Technologies, } } } } } 860-223-4710 or wwww.imagecontent.com. Ron Lund is VP sales. } } } } } } } } } } Hope this is helpful. } } } } } } } } } } Best regards, } } } } } Barbara Foster } } } } } Consortium President } } } } } Microscopy/Microscopy Education ...Educating microscopists for greater } } } } } productivity. } } } } } } } } } } } } } } } } } } } } } } I have some negatives taken on the TEM which developed } } extremely light. I } } } } } } was wondering if anyone knows any tricks on photoshop for } } generating good } } } } } } quality prints. I typically adjust the levels and condense } } the size of the } } } } } } picture, but I am still not completely happy about the quality of the } } } } } prints. } } } } } } } } } } } } Don Delaney } } } } } } } } } } } } Thomas E. Phillips, Ph.D. } } } Associate Professor of Biological Sciences } } } Director, Molecular Cytology Core Facility } } } } } } 3 Tucker Hall } } } Division of Biological Sciences } } } University of Missouri } } } Columbia, MO 65211-7400 } } } (573)-882-4712 (voice) } } } (573)-882-0123 (fax) } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
Dr Sally Stowe, Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525 http://www.anu.edu.au/EMU/home.htm
The Lawrence Hall of Science, the (nonprofit) publisher of Project MICRO's "Microscopic Explorations" middle school manual, needs micrographs for another project. The LHS is a first-rank developer of teaching materials; you can be sure that anything that you contribute will be used well. Please contact Sue Boudreau at the LHS directly: suebdoo-at-uclink4.berkeley.edu.
} "The Science Education for Public Understanding Program is developing an } activity on classification of microscopic organisms in our 7th grade life } science course. We are looking for images from the kingdoms of life, to put } on picture cards for students to sort. Each card will have two different } micrographs of the same species. } } We would like 3 representatives from each kingdom of life: plants, animals, } protists, prokaryotes/bacteria, fungi. For each, we would like ideally, } both a transmission (light or electron) and a scanning electron micrograph } (if appropriate). We are looking for examples of pathogens as well as non } pathogenic species. } } TIF images at a good publication resolution would be our preference but } JPEG } would be fine too. We would love to hear from you if you have any } images to } share with us."
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Someone has contacted me regarding having TEM done to assist in the diagnosis of Ehlers-Danlos Syndrome (an inherited disease involving collagen).
Does anyone know of an EM lab capable of doing this work? This would involve sectioning and TEM diagnosis by a pathologist.
Thank you,
John
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
You might be able to find information at at FEI Company / Philips = website ( http://www.feic.com {http://www.feic.com} ), since they have an FEG-SEM = XL series.
Ms Tang Ee Koon, Catherine Tel: 874 3216 Fax: 776 4971
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Speaking of sputter coaters, could anyone help with an old Hummer (II or = III, I think)?
I am looking for someone to come in, check it out and make it work. It = switches on, the pump makes a noise and actually draws vacuum. The = chamber looks as if it it under vacuum too. However, the vacuum gauge = does not move much past the first marker.
Does anyone know of a service group in Southern California that would be = willing to take a look at this old machine, I would really like to get it = working.
Best regards and many thanks,
Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 213 273 8026 pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
(SMTPD32-5.05) id AC19233202D0; Wed, 13 Oct 1999 20:04:25 -0500 Message-ID: {00ee01bf15e0$8629b0c0$03000004-at-gordon.rfdata.net}
} Don't forget the cost of living difference between UCLA and Indiana University. } Los Angeles is among the top 3 most expensive areas to live in this country, (it } is more expensive than Boston by 2-5%). Salaries are generally adjusted to } support a standard of living within a community. There exist many many } companies that monitor and document Cost of Living in the US and around the } world so national and global companies can adjust salaries for employees that } get moved around the states and the world. } } -geoff } } PS It does seem kinda low . . . but then I have heard of tenure track faculty } positions that start in the low $40K/year. } There are tenure track positions that start below $30K. There are enough people that want a safe small town or that don't want to leave that a university that is in a small town can pay pretty low salaries for staff.
Also what they want and what they will take in experience are not necessarily the same thing. The committees I have been on usually end up taking the one they think is best even if they don't meet the qualifications. With a salary like that they probably won't get a qualified applicant. If they don't like the top applicant they offer a salary they think he won't take. And when he refuses they make an offer to the one they want.
Sometimes jobs are advertised at low salaries so that a non qualified person that they want will be the only applicant.
Politics is not a nice game but it's the only game in town.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK 405 624-2855 GMT -6:00 www.couger.com/gcouger
PS It does seem kinda low . . . but then I have heard of tenure track faculty positions that start in the low $40K/year.
40K to start would be generous...
Ron,
Not to bust on Hoosiers, because Bobby Knight is the man, but comparing Los Angeles to South Bend, Indiana to enjoy the same standard of living with a number-cruncher program does not take into account any quality of life in these incongruent regions. Having driven through IN when I existed in KY (all bets are off there as no comparisons apply) SB and LA are worlds apart. Things like personal value sets and what kind of life someone (and their family) want to live are all more relevant than any small percentage difference that is calculated.
(I know the parties aren't making that much, it's just easier to do percents) will need to earn 85K inNot as big a difference as I thought.
This is the crux of the matter- the parties aren't making that much, peanuts, really when all is considered, so I think Mike Rock has hit the bullseye on this one. The oversupply of qualified EM and other technical people, produced by the education industry worldwide, has caused a glut in supply and our wages as a result suffer. When fewer and fewer people can be forced to do more and more, the cycle will continue.
Laura
********************************************** Laura S. Rhoads US Distributor- AM-Toffeln
L. S. Rhoads P. O. Box 554 Johnson City, NY 13790-0554
} This has been an interesting thread. However, I have one gripe. It is not } a particular software product which is giving good results with underexposed } negatives, but an algorithm which this software uses. It would be nice to } bring the discussion to this level.
you can. Ask them for their document that describes the algorithm in detail.
} The problem of making information from a negative visible under challenging } circumstances boils down to a rescaling of the pixel values by some } function, plus truncations. In addition, filters may be used, which have a } more complex dependence than simply the initial value of a given pixel, for } instance using its neighboring pixel values. I suspect the good results } with Lucis rely on a combination of rescaling and filtering. But does } anyone know what Lucis actually does? If not, one might argue that that is } bad science: using a black box without knowing what it is doing very often } comes back to haunt you. If yes, then it can in principle be done using any } software which allows sufficiently low-level processing. I'm not knocking } Lucis, but this is supposed to be a scientific forum.
Again, what Lucis does is explained in their algorithm document.
What is interesting is that what our eye sees is not always what is the total of what is in the media. This contradicts my photographic history. If the data is not in the media, it cannot be printed. This is mostly true. But by digitizing the negative and processing through Lucis or Soft-Imaging DCE, the otherwise invisible information becomes visible. This process is quite remarkable and valuable for many purposes and applications. The stereotypes and norms of the past are replaced by new technology and techniques. This is progress.
A scientific forum is one thing. But remember that what we "see" is qualitative. The point is that Lucis and DCE make images "look" good. Besides, we are not actually looking at a specimen anyway. We are looking at the effects of electron or light beams hitting and reflecting from the subject. And then there is always the Heisenberg uncertainty principle to consider.
My point is still that this software does image processing that heretofore was not possible. This is also true for Soft-Imaging's Analysis. If one dismisses the advances in computer-based signal processing, I think that is a big mistake.
} } The problem of making information from a negative visible under challenging } circumstances boils down to a rescaling of the pixel values by some } function, plus truncations. In addition, filters may be used, which have a } more complex dependence than simply the initial value of a given pixel, for } instance using its neighboring pixel values. I suspect the good results } with Lucis rely on a combination of rescaling and filtering. But does } anyone know what Lucis actually does? If not, one might argue that that is } bad science: using a black box without knowing what it is doing very often } comes back to haunt you. If yes, then it can in principle be done using any } software which allows sufficiently low-level processing. I'm not knocking } Lucis, but this is supposed to be a scientific forum. } ================= If it is patented it should be available. My quick search brought up 10,000 patents so the patent number would be a help.
I did some analysis of stream flow from x,y,z data. My method worked fine with data scanned from real plots but if I used map data I had a problem with big flat areas. The flat areas would be the same as underexposed negatives.
While I never did it, I thought that if I defined the outline of the flat area expanded the depth of the image and for every point in the flat area calculate the value base on the distance from each point upper and lower contours. Weighting the contribution of each point on the upper and lower contour lines by the inverse square of the distance between the point in question and each point on the two contour lines. This was in 286 days and I was having to page my array to disk It was also recursive so memory was a problem. so it was not practical at the time.
You should be able to extrapolate this method one step below the lowest density by using the next contour step at the centroid of the contour and using the same method. It would look funny if the contour wasn't closed.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK 405 624-2855 GMT -6:00 www.couger.com/gcouger
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK 405 624-2855 GMT -6:00 www.couger.com/gcouger
{color} {param} 0100,0100,0100 {/param} Thanks to those who responed to my question. Below is the answer I received from EDAX. Hopefully a software upgrade on my XL30 will solve the problem. I'll report back if it doesn't!
Regards
Dave
{italic} Dave,
The BSE problem that you describe is a problem with all XLs that have the
"TV/4" scan generator (very fast scan speeds that are not TV rate) and XL
software version 5.5 and earlier. Version 5.6 takes care of this in our
experience here at EDAX. We are well aware of this problem (Oxford is as
welll) but I am sure you can find many people in our organization who do
not know about this issue. The fix required in our software (and probably
in the Oxford software) is to specify a bandwidth filter setting,
otherwise it is possible the image will appear noisier than necessary or
that the edges in the image might appear smeared.
Best regards,
Bob Anderhalt
EDAX Applications Lab Manager
{nofill}
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
is there anyone out there who has tried to do postembedding labelling with samples treated with ruthenium-red before embedding. Does ruthenium-red alter the antigenicity of proteins as osmium does?
Ruthenium-red is used to keep the glycocalyx of pathogenic bacteria during the embedding procedure. Since we have no cryo-sectioning facility, does someone know how to retain the bacterial glycocalyx for further labelling studies. Thanks. Manfred
Yesterday I was trying Lucis-demo on my negatives. My negatives are dark-field images of the DNA-protein complexes with NanoGold. images is very weak and has a low contrast (except bright dot of the NG). Usually I am using Photoshop to enhance the contrast (+80-85 in Photoshop's units) and adjust gamma. The problem with these negatives is that the density of DNA much less than protein core. Adjusted conditions for DNA you send protein core out of linear part of gay-scale. Adjusting to see protein - you make DNA practically invisible. So, this is typical situation when your media do not reproduce all dynamic range of grays of the image. Usually playing with Photoshop I found some "intermediate" solution which barely acceptable. I decide such "hard" case is a very good test for Lucis. I scanned my negative as "positive transparency" in 16 bit Gray-scale mode at 1200 dpi and save it as a "tiff", then I loaded it in Lucis. Lucis-es interface itself is very simply and friendly: you have to select some particular area and play with "small"- and "big cursor" parameters (in my case the range was from 1 to 32000-something) in "pre-view" mode. When you find the best value for "small"- and "big cursor", you have a choice to apply that value to whole picture. The calculations for whole picture took all computer resources. For my picture of 50 Mb it takes 10-20 sec (double Pentium 2, 450 MHz, 486 Mb RAM). Lucis decreases the levels of gray from 16 to 8 bit. I tried approximately 100 combination for "small"- and "big cursor" value (from 1 to 32000+). The background becomes definitely smooth after the Lucis, but I did not find optimal conditions to enhance image quality. All Lucis generated images were in the frame of the Photoshop's ability and do not show more/better details at least in my hands. The conclusion is: if something on the original picture is poorly visible or has low contrast, after Lucis treatment it is poorly visible too. This conclusion based only on my very limited experience with Lucis. May be for another application this software will work better. I am sorry, Lucis.
And I am, also, impressed by price. The program has very limited set of functions. You are not able even crop the picture to speed the calculation. In my point of view even "discount price" of 1000+something$$ are not reasonable. This program may be useful in some very special case which cover, may be only 0.1% of EM needs.
Sincerely yours,
_________________________________
Sergey Ryazantsev Ph. D. UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
"I am also very impressed by the filter function DCE (differential contrast enhancement) that the German software called "analySIS" has included. It also does much more than a simple "contrast/brightness" or even "gamma" adjustment. And it's also working fantastic with color images! A sample can be viewed on their website www.soft-imaging.de ...."
Kind regards!
Martin Bartels AG Plant Ecology C.v.O. University Oldenburg email: bartels-at-uni-oldenburg.de tel. (+49)441 798-3436
Let us refer to the "coupler" as "the stuff the cell is in." It is unclear from your message whether you are dealing with a mounted, under a cover glass, or just on the slide cell. Mounting medium, as opposed to the oil used with oil lenses, are two different things.
} From BIOLOGICAL MICROTECHNIQUE by J. B. Sanderson from the Royal Microscopy Handbook Series.
"Mounting media differ according to whether they are used to mount stained or unstained tissues. In the former case, it is important that the medium has a RI as close to that of glass as possible so that the detail demonstrated by the stain can be seen clearly without interference. Where the specimen of tissue is unstained, the RI of the mounting medium should differ from that of the tissue as much as possible. In this way the medium is used to enhance the tissue detail that would otherwise be lost."
You are looking at difference. When the RI is the same as the tissue it basically becomes invisible, and you can't get much less contrast than that. AS the RI of the "stuff the cell is in" moves away from the cells, either up or down, the contrast increases. Loss of detail and contrast are not the same thing. You might be mixing the two up.
I hope this helps.
Shalom from Jerusalem, Azriel Gorski
At 08:53 13-10-99 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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It is my understanding that the Lucis software is the commercialization of the "Differential Hysteresis" software developed by Klaus-Ruediger Peters at the University of Connecticut. Klaus has presented talks on this at MSA meetings in previous years. A website describing this work is: http://www.cbit.uchc.edu/indiv/software_docs/dhp.html
Klaus's original software was extremely processing-intensive. When first written, it required something like a Silicon Graphics workstation. As I understand the algorithm, it revolves a series of vectors through each pixel in the image. Searching out along each of these vectors, the intensity values are compared with a pair of bracketed thresholds that are changed in kind of quantum steps. It is a quite sophisticated algorithm and I have been impressed with the demonstrations I have seen. I have always been impressed with Klaus's grasp of the theoretical aspects of imaging and my perception is that this really is a new and novel algorithm.
My understanding is that the Lucis software has been refined to permit its use on a PC. The demonstrations I have seen were reasonably fast -- much faster than I would have expected from my first exposure to the algorithm.
We had this idea that it would be very convenient to use the SEM just to have a quick look at sections on the grid - in the long run as a tool for observing the result of the immuno gold labelling (in addition to the TEM.)
As a trial we used osmicated but not uranylacetate/lead citrate treated sections (which are also immuno labelled with 10nm gold).
The grid was positioned and stuck on top of a hole drilled in a carbon rod and mounted on a stub.
Although we did not really expect to see the gold particles (because they were not silver enhanced) we were quite disappointed that neither in the backscattered electron mode nor in the secondary electron mode did we see more than a vague outline of the cells.
Is there a way to see some details of the fine structure (mitochondrias, nuclei..) in ultrathin sections in the SEM?
Your expertise is as usual highly appreciated.
Thank you very much.
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK ++44(0)181 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
Once again, I can't help but say that some of us are content with our salaries, paying mortgages, and going to Disney world. You've got to consider the source. Universities don't pay high salaries unless you are a mathematician or an engineer. Science pays by knowing your professional contribution has furthered our understanding of what makes this world go round. If you want the big buck boys and girls, consider big business, where you will have to watch your back and never see your families. Oh, but you'll have a smashing house. Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
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I work at Purdue University in West Lafayette, Indiana. That OTHER university in Bloomington, Indiana has cost-of -living similar to ours. I can't imagine expecting a PhD level person to accept this position at the stated salary unless it was the last option. It may be appropriate for a MS level but I would assume that the individual would need to learn some of the required techniques on the job.
That's a pretty hard job description to fill at any salary but I wish them luck finding someone. I think it is great that they recognize the importance of microscopy to the degree that they have created this new position (it is new...personal communication). Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
I agree, this is a low salary even here in Dallas, where the cost of living is much lower than in Cal. I hope the employer in Indiana has someone specifically in line for the job or that the cost of living in Indiana is still lower than Dallas. I guess if your an NIH post-doc making 22-24K a year, this could look promising. However, post-docs haven't usually demonstrated excellence in so many areas of microscopy.
} } } It is unbelievable: such duties for $37K?! There, at UCLA $37K is a level } of very moderate post-doc. To be equally perfect in the "light, confocal, } fluorescence, transmission, and scanning electron microscopes" just } impossible. After 15 years working with EM I would say : I am moderately } "perfect" in some very narrow TEM area. This is reason I will not be a } successful candidate for that position as well as for many others. } } Sergey } } } Date: Mon, 11 Oct 1999 18:37:28 -0500 } } From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.microscopy.com} } } Subject: LM,TEM,SEM Job Announcemen } } 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.Co m } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm l } } -----------------------------------------------------------------------
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Dear Nelson and all,
Apparently, several people have confused my with the person offering the job. I am not that person and I have no job to offer. The offer was for a job somewhere in Indiana. My comment was that the money seemed a bit low to me for a PhD with expertice in so many areas of microscopy and who would be expected to run the lab, teach, and do independant research.
Nelson, I don't know where you are, but by your e-mail address it looks as though you are outside the US. I know many students that come here from all over the world who think they are going to have an easy time with living expences only to find out that they didn't figure in all the little things like taxes, car(a must in Dallas), all the little things that aren't covered by tuition, etc. I know people think Americans are spoiled and maybe we are, but the position offered would typically go for a fair amount more even in Dallas, where the cost of living is substantially lower than in Callifornia. They said they wanted a PhD with "expertise" in most of the microscopic techniques used in biology. This person most likely will be expected to run the lab-supervise personnel, keep the books, maintain the equipment, teach, maybe even run up sample for other investigators and do independant research. Sounds like about a 60-80 hour a week job, more when grants are due.
On Wed, 13 Oct 1999, Nelson Fava wrote:
} Dear Miss Pawlowski, } } I would like to say that I, an electron microprobe specialist, working and doing } maintenance in a CAMECA SX50 (serial #359), four WDS, one EDS, BSE, SE and X-Ray } imaging generation apparatus, during seven years, would appreciate to work in } Dallas for 30K/year. Besides, I do the same job with: X-Ray powder diffraction } equipment, Termo-Differential Analyzer, ICP-AES equipent and a MAT 250 hot } cathode mass spectrometer. } } Please find attached my resume, } } Thanks in advance, } } Sincerely, } } Nelson Fava. } } } } } Karen S Pawlowski wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } I agree, this is a low salary even here in Dallas, where the cost of } } living is much lower than in Cal. I hope the employer in Indiana has } } someone specifically in line for the job or that the cost of living in } } Indiana is still lower than Dallas. I guess if your an NIH post-doc } } making 22-24K a year, this could look promising. However, post-docs } } haven't usually demonstrated excellence in so many areas of microscopy. } } } } Just my two cents. } } } } Karen Pawlowski } } Sr. Res. Assoc./UT Southwesten Med. Ctr. } } PhD Student/UT Dallas } } Dallas, TX } } } } On Tue, 12 Oct 1999, Sergey Ryazantsev wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } It is unbelievable: such duties for $37K?! There, at UCLA $37K is a level } } } of very moderate post-doc. To be equally perfect in the "light, confocal, } } } fluorescence, transmission, and scanning electron microscopes" just } } } impossible. After 15 years working with EM I would say : I am moderately } } } "perfect" in some very narrow TEM area. This is reason I will not be a } } } successful candidate for that position as well as for many others. } } } } } } Sergey } } } } } } } Date: Mon, 11 Oct 1999 18:37:28 -0500 } } } } From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.microscopy.com} } } } } Subject: LM,TEM,SEM Job Announcemen } } } } 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 } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Direct all inquiries to the address below. } } } } } } } } Nestor } } } } } } } } ------------------------------------------------ } } } } } } } } } } } } } } From: Rhea Freeman {freemanr-at-bio.indiana.edu} } } } } } } Subject: LM,TEM,SEM Job Announcement } } } } } } } } } } } } } } } } } } } } } } } } } JOB ANNOUNCEMENT } } } } } **************** } } } } } The Indiana Molecular Biology Institute at Indiana University is seeking a } } } } } Ph.D level scientist with demonstrated excellence in microscopy to direct } } } } } the Molecular Biology Microscopy Facilities, which include light, confocal } } } } } fluorescence, transmission, and scanning electron microscopes. The } } } } } position includes overall management of the facilities, user training, and } } } } } user supervision. We seek a candidate who will be an active participant } } } } } with Institute faculty in the development of our microscopy capabilities } } } } } and training programs. In addition, pursuit of independent and } } } } } collaborative research will be strongly encouraged. Salary: $37,000. } } } } } } } } } } Candidates should send a resume and have three letters of reference } } } } } sent to Dr. Rudolf A. Raff, Director, Indiana Molecular Biology Institute } } } } } Indiana University, Jordan Hall, 1001 E. Third St., Bloomington, } } } } } IN 47405. Deadline for application: Nov. 1, 1999. Indiana University } } } is an } } } } } Equal Opportunity/Affirmative Action Employer. } } } } } } } } } } Additional information is available at } } } } } http://www.bio.indiana.edu/research/imcb/MFM_Ad.html } } } } } } } } } } -------------------------------------------------------------------------- } } } ----- } } } } } } } } } } Rhea Freeman Ph. 812-855-4183 } } } } } Administrative Assistant Fax 812-855-6082 } } } } } Indiana Molecular Biology Institute } } } } } Jordan Hall 322A } } } } } Indiana University } } } } } Bloomington, IN 47405 } } } } } } } } } } } } } } } } } } } } } } } } _________________________________ } } } } } } Sergey Ryazantsev Ph. D. } } } UCLA School of Medicine } } } Department of Biological Chemistry } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } Phone: (310) 825-1144 } } } FAX (departmental): (310) 206-5272 } } } mailto:sryazant-at-ucla.edu } } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } } } } }
Our laboratory has an Leica/Reichert MEF4 metallograph fitted with a Ludl moveable stage. The autofocus does not work with the 5x objective. Ludl tells me that this is a known problem with their stage and the MEF4 microscope and the only way to overcome this problem is by changing the image acquisition software.
Is there anyone else who has this microscope/stage combination and is experiencing similar autofocus problems? I would like to know how you solved the problem. I am currently using ImagePro Plus software, but am willing to consider other software solutions.
Hello all: I am hoping one of the list members can help with this issue. Is there a rule of thumb or more specific guidelines regarding energy delivered to living cells when imaged with a LSCM compared to a standard fluorescent microscope.
Ramin Rahbari PARKE-DAVIS Pharmaceutical Research Pathology and Experimental Toxicology 2800 Plymouth Road Ann Arbor, MI 48105 Voice (734) 622-3383 Fax (734) 622-5001 Ramin.Rahbari-at-WL.COM
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We have had some experience using ruthenium hexammine tetroxide FOLLOWING enbloc (diffusion) labeling for EM using decorin (a small lucine-rich proteoglycan which peridically "decorates" collagen fibrils in skin and other tissues) antibody. We expected that decorin would be leached from tissue during the prolonged incubations in immunocytochemical solutions and rinses, but instead it was retained and successfully labeled. Please see Keene et al., J. Histochem. Cytochem. 46, page 215 (1998). I am not sure that this techique will work on bacteria, but it may be worth a try.
I hope this helps,
Doug ---------------------- Douglas R. Keene Shriners Hospital Microscopy Facility 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 503-221-3434 DRK-at-shcc.org
Hi everyone, Is there a protocol of beta-gal staining for vibratome cut thick section? The enzyme activity in mouse organ is always hard to detect. The blue color is in the rim of paraffin tissue sections. I'm thinking fresh unfixed 100-200 microns section will probably penetrate easier. Hope I will get some hint in some points. Thanks.
Dorothy Zhang Harvard School of public Health Building 2, CVLAB 677 Huntington Ave, Boston, MA 02115 Phone# 617-432-6981 Fax# 617-432-2980
} It is my understanding that the Lucis software is the } commercialization} of the "Differential Hysteresis" } software developed by Klaus-Ruediger ...
More info and nice A-B comparisons can be found at:
http://www.imagecontent.com/lucis/default.htm
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Email: canew-at-jps.net Name: C. Newhouse School: Lowell HS
Question: I am interested in finding a text or literature on slide preparation & staining techniques (& preparation of stains including Congo red, Safranin, Chrystal Violet, etc.)
Claudia: You have re-discovered that the SEM gives largely topographic contrast and sections are flat. In backscattered mode you get atomic number contrast, but there is not enough atomic number difference within biological section. The gold, would be near the limit of resolution, certainly under these bad viewing conditions and in BS mode. You could do a little better if you looked at the block-face (with a whiff of carbon coating) and better still, if that block-face was etched.
In the case of the thin sections, matters are much worse because most of the electrons pass through - and mostly these will never contact either the 2ndary or the BS detectors. If you had a photomultiplier under the section, this would be rather like a STEM in some ways, and in that configuration you could get somewhat better imaging, but you would require about 500nm sections to get 'half-decent" results. I don't think that your present technique will yield useful results. However, you would optimize the imaging by: Use the lowest kV that still gives reasonable emission Use fairly high emission - a bit over 100 uA Spread the condenser Increase the photomultiplier (or BS) contrast until the image appears just a little grainy. Adjust brightness with that potentiometer.
Its not likely to be useful, but its a nice learning experience. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, October 14, 1999 10:25 PM, Claudia Hayward-Costa [SMTP:LS_S562-at-crystal.kingston.ac.uk] wrote: } } Dear List, } } We had this idea that it would be very convenient to use the SEM } just to have a quick look at sections on the grid - in the long run as } a tool for observing the result of the immuno gold labelling (in } addition to the TEM.) } } As a trial we used osmicated but not uranylacetate/lead citrate } treated sections (which are also immuno labelled with 10nm gold). } } The grid was positioned and stuck on top of a hole drilled in a } carbon rod and mounted on a stub. } } Although we did not really expect to see the gold particles } (because they were not silver enhanced) we were quite } disappointed that neither in the backscattered electron mode nor in } the secondary electron mode did we see more than a vague outline } of the cells. } } Is there a way to see some details of the fine structure } (mitochondrias, nuclei..) in ultrathin sections in the SEM? } } Your expertise is as usual highly appreciated. } } Thank you very much. } } Claudia } } Dr. C. Hayward-Costa } School of Life Sciences } Kingston University } Penrhyn Road, Kingston upon Thames } Surrey KT1 2EE, UK } ++44(0)181 547 2000 x 2240 } Email: c.hayward-at-kingston.ac.uk }
The latest edition of "Precision" from Hitachi describes a simple method of getting "STEM" images in a SEM. If you get in touch with them I am sure they will send a copy.
Paul Ansell ( Sales Manager ) may be contacted at:-
paula-at-nissei-eu.com
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie http://www2.tcd.ie/Electron_Microscope/emu/home.htm
-----Original Message----- } From: Claudia Hayward-Costa [SMTP:LS_S562-at-crystal.kingston.ac.uk] Sent: Thursday, October 14, 1999 1:25 PM To: Microscopy-at-sparc5.microscopy.com
Dear List,
We had this idea that it would be very convenient to use the SEM just to have a quick look at sections on the grid - in the long run as a tool for observing the result of the immuno gold labelling (in addition to the TEM.)
As a trial we used osmicated but not uranylacetate/lead citrate treated sections (which are also immuno labelled with 10nm gold).
The grid was positioned and stuck on top of a hole drilled in a carbon rod and mounted on a stub.
Although we did not really expect to see the gold particles (because they were not silver enhanced) we were quite disappointed that neither in the backscattered electron mode nor in the secondary electron mode did we see more than a vague outline of the cells.
Is there a way to see some details of the fine structure (mitochondrias, nuclei..) in ultrathin sections in the SEM?
Your expertise is as usual highly appreciated.
Thank you very much.
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK ++44(0)181 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
hey all, I am searching for a preparation technique for humic acids for TEM. Are there any experiences with negative staining methods? with regards wolfgang wiehe
Richards, RG & Ap Gwynn, I (1995) "Backscattered electron imaging of the undersurface of resin-embedded cells by field-emission scanning electron microscopy" J Microscopy 177 (1) 43-52.
I think they looked at blocks rather than sections.
Dave
On Thu, 14 Oct 1999 13:24:44 +0100 Claudia Hayward-Costa {LS_S562-at-crystal.kingston.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear List, } } We had this idea that it would be very convenient to use the SEM } just to have a quick look at sections on the grid - in the long run as } a tool for observing the result of the immuno gold labelling (in } addition to the TEM.) } } As a trial we used osmicated but not uranylacetate/lead citrate } treated sections (which are also immuno labelled with 10nm gold). } } The grid was positioned and stuck on top of a hole drilled in a } carbon rod and mounted on a stub. } } Although we did not really expect to see the gold particles } (because they were not silver enhanced) we were quite } disappointed that neither in the backscattered electron mode nor in } the secondary electron mode did we see more than a vague outline } of the cells. } } Is there a way to see some details of the fine structure } (mitochondrias, nuclei..) in ultrathin sections in the SEM? } } Your expertise is as usual highly appreciated. } } Thank you very much. } } Claudia } } Dr. C. Hayward-Costa } School of Life Sciences } Kingston University } Penrhyn Road, Kingston upon Thames } Surrey KT1 2EE, UK } ++44(0)181 547 2000 x 2240 } Email: c.hayward-at-kingston.ac.uk }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
We have a digital camera Sony make , Model: MVC -FD7. We would like to use the same for taking microphotographs by attaching it to the optical microscope. Somehow we want to use this camera for taking microphotographs, we have already tried a lot but could not succeed. Could you please help/guide us on using this Sony digital (MVC-FD7) camera for taking microphotographs?.
Regards,
Girish Shejale Sr. Executive Life Extension Services. Thermax Babcock & Wilcox Limited Pune, India.
---------------------- Forwarded by Girish Shejale/TBWL on 10/15/99 08:58 AM ---------------------------
"Jeff Stewart" {jeff-at-metallography.com} on 10/14/99 04:48:50 PM
To: Girish Shejale/TBWL cc:
Thank you Tracey. My sentiments exactly.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75235-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
} } } "Tracey M. Pepper" {tpepper-at-iastate.edu} - 10/14/99 8:44 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Once again, I can't help but say that some of us are content with our salaries, paying mortgages, and going to Disney world. You've got to consider the source. Universities don't pay high salaries unless you are a mathematician or an engineer. Science pays by knowing = your professional contribution has furthered our understanding of what makes = this world go round. If you want the big buck boys and girls, consider big business, where you will have to watch your back and never see your = families. Oh, but you'll have a smashing house. Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
We have not been able to embed Golg-Cox brain tissue in JB4. The JB4 solidifies, but the tissue within it does not. We have done it with and without polyethylene glycol. We have infiltrated the tissue with A+C for varying lengths of time. We have tried dehydrating the tissue in 100% ethanol prior to infiltration. We just purchased new JB4, so the solutions are not old. Any suggestions would be greatly appreciated.
Sorry, but I have to disagree with your assessment of working for industry. I do not have to watch my back and I see my family more than I would if working for academia because I have the money to fly to visit them. When I saw the announcement, my reaction was "tell me again why I went into industry". And salary doesn't even begin to cover the plus side of my work, who I work for, and with whom I work.
Damian Neuberger
Once again, I can't help but say that some of us are content with our salaries, paying mortgages, and going to Disney world. You've got to consider the source. Universities don't pay high salaries unless you are a mathematician or an engineer. Science pays by knowing your professional contribution has furthered our understanding of what makes this world go round. If you want the big buck boys and girls, consider big business, where you will have to watch your back and never see your families. Oh, but you'll have a smashing house. Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
I am looking for advice and experience from people who used SEM as an analytical tool i.e. EDS and WDX analysis of geological and metallurgical samples. We are evaluating the pros and cons of a E- probe vs SEM with EDS-WDX possibilities
Our situation : - not a high throughput - main samples types are geological and metallurgical - main uses chemical determinations
The questions are then : What are the pros and cons of using a well equiped SEM vs a E- probe? How good/bad SEM are probes ? How good/bad probe are SEM ?
Some of our understanding : - probe columns are optimized for analysis while SEM for imaging - larger current possibilities on probes - multiple spectro on probe and simultanous WDX analyses (sequetial and a SEM) - light elements ---} probe - SEM are more versatile tools - probe are more expensive$
Please answer personnally and I will post a wrap-up at the end. Thanks for your help,
..by the way, Vanderbilt needs an EM person. Although I don't know whether you would like the Southern surroundings, it is a lower pressure environment than UCSF, and there is a local art scene. Even if you arent interested, you could pass this opportunity around to anyone you know who might be qualified.
Contact directly:
Peter Kolodziej, Ph.D. 823 Light Hall Howard Hughes Medical Institute Vanderbilt University Medical Center Nashville TN 37232-0295
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} We have not been able to embed Golg-Cox brain tissue in JB4. The JB4 } solidifies, but the tissue within it does not. We have done it with and } without polyethylene glycol. We have infiltrated the tissue with A+C } for varying lengths of time. We have tried dehydrating the tissue in } 100% ethanol prior to infiltration. We just purchased new JB4, so the } solutions are not old. Any suggestions would be greatly appreciated.
We gave up on JB-4 for this years ago and developed a soft araldite (later soft Epon) method which has worked well.
} -geoff, } } PS It does seem kinda low . . . but then I have heard of tenure track faculty } positions that start in the low $40K/year.
40K to start would be generous...
} Ron, }
Not to bust on Hoosiers, because Bobby Knight is the man, but comparing Los Angeles to South Bend, Indiana to enjoy the same standard of living with a number-cruncher program does not take into account any quality of life in these incongruent regions. Having driven through IN when I existed in KY (all bets are off there as no comparisons apply) SB and LA are worlds apart. Things like personal value sets and what kind of life someone (and their family) want to live are all more relevant than any small percentage difference that is calculated.
} } (I know the parties aren't } making that much, it's just easier to do percents) } will need to earn 85K inNot as big a difference as I thought. }
This is the crux of the matter- the parties aren't making that much, peanuts, really when all is considered, so I think Mike Rock has hit the bullseye on this one. The oversupply of qualified EM and other technical people, produced by the education industry worldwide, has caused a glut in supply and our wages as a result suffer. When fewer and fewer people can be forced to do more and more, the cycle will continue.
Laura ********************************************** Laura S. Rhoads US Distributor- AM-Toffeln
L. S. Rhoads P. O. Box 554 Johnson City, NY 13790-0554
The standard is Dr. Freida Carson's book, the title of which just flew out of my head. It is available through the ASCP press and web sites like Amazon.com, as it is in print. All the stains you mentioned are in there, and many more. You can contact me off list for more information on the subject. I am a registered histotech, and tissue processing, slide preparation and staining are, quite literally, my life. Wanda Shotsberger Harris Methodist Hospital Fort Worth TX ---------- } From: "canew-at-jps.net"-at-Sparc5.Microscopy.Com To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Email: canew-at-jps.net Name: C. Newhouse School: Lowell HS
Question: I am interested in finding a text or literature on slide preparation & staining techniques (& preparation of stains including Congo red, Safranin, Chrystal Violet, etc.)
} Email: canew-at-jps.net } Name: C. Newhouse } School: Lowell HS } } Question: I am interested in finding a text or literature on } slide preparation & staining techniques } (& preparation of stains including Congo red, } Safranin, Chrystal Violet, etc.) } } ---------------------------------------------------------------------------
Mr. or Ms. Newhouse -
It isn't easy to answer your question without a bit more information. Are you writing a report about existing histological preparations, or are you considering making slides yourself? There are two books in the MICRO bibliography (URL below) that can help you:"Exploring with the Microscope" by Nachtigall, and "The Microscope on a Budget" by Stevens. And the Carolina Biological catalog lists "Laboratory Manual of Histology" by Pappas as #D8-45-5902. I haven't seen the Pappas book, but it probably has the detail that you want. But to quote Nachtigall, "I do not recommend that you make your own permanent slides, because the process is complicated, takes a long time, and the outcome is seldom what you expected." If you want to try anyway, you'll find instructions for making hand-cut hematoxylin & eosin sections of liver in Nachtigall - which will introduce you to the basic process.
If you'd like a microscopist-advisor in San Francisco, I can find you one.
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Dorothy I have not done much beta-gal localization; but have done lots of cerium based histochemistry. I know of two things which will improve the reaction with lightly fixed tissues. 1) Use a 30 minute preincubation step in 37C water bath which includes all incubation components except the SUBSTRATE;then incubate for 30 minutes in the complete reaction medium wgich contains the SUBSTRATE; 2) include 0.0001-0.0002% Triton X-100 in both the preincubation and complete reaction medium [make a 1% solution of Triton X-100 in deionized water and add 1-2 drops of this solution to each 10 ml of reaction medium. You probably will not get improved penetration with fresh unfixed tissue unless you do a freeze-thaw and that may result in structural degradation. Ann Ellis College of Medicine/College of Veterinary Medicine University of Florida Gainesville, FL FAX (352)846-2231
A sincere thank you to all of you who took the time to send me advice, literature sources, encouragement and helpful comments. What a great place this list is!
Claudia Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK ++44(0)181 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
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We demo'd the new Spot RT camera last week. It was really quite impressive. The specifications are on the web at: http://www.diaginc.com/spotrts.htm It is a supstantial improvement over the older Spot and Spot-2 cameras in that focusing is much closer to real-time and on the whole image...in B&W or color. Capture is also quite fast. There is substantial ability to control all camera features regarding exposure, gain, color balance, etc. It also is available for either MAC or PC. It is also a true 12-bit camera with resolution at 1520 x 1080 optical resolution. Actual capture speed depends on computer, video card, sample brightness, color or monochrome, and final resolution but can reach 12 frames/sec on color and 19 frames/sec for monochrome. Price ranges from about $7500 to $12500 without adapter tubes for specific microscopes and a computer. Although venders are not likely to have the camera at the moment (the company rep did our demo), they should get them soon. It is certainly worth the wait to test it out before making a purchase of another camera. I have also demo'd the CoolSnap. It is a very nice camera ..possibly the best at the lower end of the cost spectrum. (approx. $6000+ adapter tubes, etc.) Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
Message-ID: {3807243E.60B307E9-at-nki.nl}
After installation and working a lot with my new EDS systems (Imix PC) from PGT, I am now preparing publications. And - surprise, surprise, - I have found out that I cannot print good quality pictures (on high quality printer which is not connected to our network)!
The file format of stored images, RAS with overlays, cannot be read by other programs. Of course, some of them can read RAS, but overlays (with the most important information) are lost.
The only advice I've got from PGT was to capture images from the monitor. But then I will have files with much lower resolution than initial ones, and this is certainly bad for high quality printing.
All (if any) advises will be highly appreciated!!!
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Aaron: The methacrylate media generally do not work well in the presence of metals. This is particularly true of osmium, and in my experience has included the silver and gold-toned stains as well. Certainly your comment about the bulk plastic polymerizing while the plastic immediately surrounding the stained neurons not polymerizing would seem to corroborate my earlier anecdotal experience. I too think that a soft epoxy (araldite is an excellent choice) would be a preferable alternative.
Roger Moretz Dept of Toxicology
} -----Original Message----- } From: Aaron R. Best [SMTP:a_best-at-ou.edu] } Sent: Friday, October 15, 1999 11:26 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM-Embedding Golgi-Cox in JB4 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have not been able to embed Golg-Cox brain tissue in JB4. The JB4 } solidifies, but the tissue within it does not. We have done it with and } without polyethylene glycol. We have infiltrated the tissue with A+C } for varying lengths of time. We have tried dehydrating the tissue in } 100% ethanol prior to infiltration. We just purchased new JB4, so the } solutions are not old. Any suggestions would be greatly appreciated.
Are you sure that you don't have some sort of export function available? Most units have something aavailable, but they do not always export annotations or scale bars.
What sort of pixel resolution are you using for your images? Are you sure that an image capture would not do the job? We usually run our screens at 1280 and sometimes 1600 pixels across on a 19" or larger monitor. That provides a lot of pixels for a screen capture. Remember that you can use PrtScrn to capture the entire desktop area or Alt-PrtScrn to capture just the active window to the clipboard for pasting into a word processor or an image processing application. images will get captured at the current screen resolution and color depth (i.e., 8-bit/256 color, 16-bit/64K color, or 24-bit/true color).
You don't say what kind of high quality printer you have available, so I will have to assume a few things.
First, let me suppose you are using a 1200 dpi laser printer. It take multiple printer pixels to fairly represent a single image pixel in gray scale. An array of 12x12 printer pixels could represent 144 shades of grade from an image pixel, which should be enough to appear to be continuous gray tones to the human eye. Even an 8x8 array may be adequate. That means a 1600 pixel image would need to be printed out at 16 inches wide on a 1200 dpi printer to show all pixels at continuous gray scale. (16 inches = 1600 image pixels * (12 printer pixels/1 image pixel) / 1200 printer pixels per inch) I doubt that you really want to print that large. To print smaller, you must sacrifice either gray scale depth or the number of pixels in an image.
Second, let me assume you are using a dye-sub printer at 300 dpi. In that case, each printer pixel can pretty well represent the full range of gray scale for a single image pixel. under these circumstances, a 1600 pixel image can be rendered at full resolution in 5.3 inches.
Practically, what we do is to collect our images at 1024 pixels across. We can thus get pretty close to full resolution on an 8-1/2 x 11-inch page. If we need to zoom in on some areas of interest, we simply do so with the microscope and take additional images.
Now, if you cannot capture 1024-pixel images from your monitor, I suggest it is time to upgrade your video card and monitor to be able to display 1280 pixels across. Good video cards are available for well under $100. A 19-inch monitor can be had for well under $1000. If your PC conforms to standards, it should be a small matter to make the upgrade.
Hope this helps.
Warren S.
At 10:34 AM 10/18/1999 -0500, you wrote: } } After installation and working a lot with my new EDS systems (Imix PC) } from PGT, I am now preparing publications. And - surprise, surprise, - } I have found out that I cannot print good quality pictures (on } high quality printer which is not connected to our network)! } } The file format of stored images, RAS with overlays, cannot be read by } other programs. Of course, some of them can read RAS, but overlays } (with the most important information) are lost. } } The only advice I've got from PGT was to capture images from the monitor. } But then I will have files with much lower resolution than initial ones, } and this is certainly bad for high quality printing. } } All (if any) advises will be highly appreciated!!! } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager
There isn't any need to reply to this message to be removed from our mailing list. This is a 1 time mailing, you will not be contacted again. Thank You for your patience.
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} Is there anyone out there that uses large quantities of electron image film? } If so, what kind of dessicant or pre-pump procedure do you use to dessicate } film quickly? } We go through quite a bit of film (Kodak ISO-163 for EM) and are looking for } an 'evironmentally friendly' film dessicant. Currently, we are using P2O5 } powder in a vessel that we place in our cylindrical vacuum pump, in which we } dessicate our film prior to loading the cassette into the electron } microscope. I've ordered recyclable dessicant in a canister to try out, but } wanted to see how others are dealing with this aspect of microscopy. } } Look forward to hearing from you all :) }
Dear Figan, We have (at least) two systems; one uses P2O5, and the other uses Mg(ClO4)2. Each works well. I'm not sure how unfriendly the amount of P2O5 you use would be to the environment. That depends on what the total use is and where the P ends up. If it's in a lake, that's bad, but if it's in fertilizer made from treated sewage, that's good. Also, if the P2O5 is a minute fraction of the P in the waste water stream, there might not be a measurable effect from your addition, but the environmental cost of producing an alternative dissicant could be measurable. Yours, Bill Tivol
Dear Volodya, Try InfanView (mailto:e9227474-at-student.tuwien.ac.at - I did not try this address). It is pretty good viewer and it may open RAS files. I did not try InfanView to print out, but you may try to save your image in some suitable format like "bmp" or "tiff" and print later from Photoshop for instance. Actually, I am really happy with that "viewer". It is very fast, recognizes most popular image formats, "slide show" option, direct viewing the Directory content etc. After a hard search of the Internet, I choose this program as a "default" viewer. InfanView is freeware, so, I have no any financial as well as other interest in this product. Good luck, Sergey.
} Date: Mon, 18 Oct 1999 10:34:22 -0500 } From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} } Subject: Imix PC pictures } To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} } X-Mailer: Internet Mail Service (5.5.2448.0) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I also am interested in a camera setup for dual-label fluorescence. Originally I was thinking of a color CCD camera but now am wondering whether it would be better to get a monochrome with an automated filter wheel. The trick for me is that, while my specimens can be labeled fairly brightly, they are particulate, suspended in saline wet mounts, so there is motion due to convection. With a video camera using brightfield, even if I wait for slow-moving particles I sometimes see "steps" in the edges of the particles, indicating that 1/30 sec is barely fast enough. So a cooled camera won't help me; I need sensitivity instead. Can anyone recommend a *sensitive* color or monochrome camera? Video is OK but a 10 or 12 bit digital one that's not extremely expensive would be better. At least several frames per second for focusing would be important. Any comments on the DVC 1300 cameras?
Can you recommend an automated filter wheel & control software? I'm not familiar with this at all. I have an Olympus BX50 scope.
Also, does anyone have suggestions for something like xanthan gum to add to the saline suspending medium to slow down convection? My particles are not alive but are sensitive to temperature and to osmotic changes.
Thank you! Richard Thrift Richard_Thrift-at-SkyePharma.com
} } } Michal Opas {m.opas-at-UTORONTO.CA} 10/18/99 11:39:27 AM } } } Dear all,
I realize that a topic of CCDs has been discussed here a few times. While pros and cons of CCD use were discussed, I am not sure if any recommendation as to what to buy it terms hardware was put forward. I am not sure, furthermore, if any guidelines for "the masses" have been established in terms of how to tackle setting up a CCD-based fluorescence detection system that would replace photoprocessing.
So there we go:
I would like to set up a "departmental" microscopical "facility" devoted to fluorescent immunolocalization. I predict just a few ( { 10) users. We have a Photomicroscope III that has been used for that purpose. I would like to equip this microscope with hardware/software such as to dispense with photoprocessing. Your advice shall be most welcome.
Cheers Michal
Dr. Michal Opas University of Toronto 1 King's College Circle Medical Sciences Building, rm 6342 Toronto, Ontario, M5S 1A8 Canada -------------- phone: (416) 978-8947 fax: (416) 978-3954 e-mail: m.opas-at-utoronto.ca
You could just abandon the dessicant and rely on the rotary pump....works for us on a number of systems, and I'd be doubtful of how much extra advantage you get from the dessicant, especially in a short time. Sally Stowe
Seiler,Figen wrote:
} Is there anyone out there that uses large quantities of electron image film? } If so, what kind of dessicant or pre-pump procedure do you use to dessicate } film quickly? } We go through quite a bit of film (Kodak ISO-163 for EM) and are looking for } an 'evironmentally friendly' film dessicant. Currently, we are using P2O5 } powder in a vessel that we place in our cylindrical vacuum pump, in which we } dessicate our film prior to loading the cassette into the electron } microscope. I've ordered recyclable dessicant in a canister to try out, but } wanted to see how others are dealing with this aspect of microscopy. } } Look forward to hearing from you all :) }
Dr Sally Stowe, Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525 http://www.anu.edu.au/EMU/home.htm
our software, analySIS, does read RAS images (SUN Raster Images). It can then save them in a large number of other formats. If the number of images is not too large, I could try to convert them for you. I can't guarantee that it works, but maybe worth a shot. Let me know if you are interested. Since we are not in the business of converting files, we are not going to charge you for the conversion, but we will limit the time we spend on this.
Note to everybody: I am just trying to help Dr. Dusevich with his images. Please do not send me any images for conversion without discussing that with me first. I will not be responsible for images sent to me without prior discussion.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Dusevich, Vladimir[SMTP:DUSEVICHV-at-UMKC.EDU] } Sent: Monday, October 18, 1999 9:34:22 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Imix PC pictures } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
After installation and working a lot with my new EDS systems (Imix PC) from PGT, I am now preparing publications. And - surprise, surprise, - I have found out that I cannot print good quality pictures (on high quality printer which is not connected to our network)!
The file format of stored images, RAS with overlays, cannot be read by other programs. Of course, some of them can read RAS, but overlays (with the most important information) are lost.
The only advice I've got from PGT was to capture images from the monitor. But then I will have files with much lower resolution than initial ones, and this is certainly bad for high quality printing.
All (if any) advises will be highly appreciated!!!
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
The Lawrence Hall of Science, the (nonprofit) publisher of Project MICRO's "Microscopic Explorations" middle school manual, needs micrographs for another project. The LHS is a first-rank developer of teaching materials; you can be sure that anything that you contribute will be used well. Please contact Sue Boudreau at the LHS directly: suebdoo-at-uclink4.berkeley.edu
} "The Science Education for Public Understanding Program is developing an } activity on classification of microscopic organisms in our 7th grade life } science course. We are looking for images from the kingdoms of life, to put } on picture cards for students to sort. Each card will have two different } micrographs of the same species. } } We would like 3 representatives from each kingdom of life: plants, animals, } protists, prokaryotes/bacteria, fungi. For each, we would like ideally, } both a transmission (light or electron) and a scanning electron micrograph } (if appropriate). We are looking for examples of pathogens as well as non } pathogenic species. } } TIF images at a good publication resolution would be our preference but } JPEG would be fine too. We would love to hear from you if you have any } images to share with us."
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Do you know what the following people have in common?
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They are a few of the 50 outstanding scientists that have received the prestigious Microscopy Society of America Distinguished Scientist Awards!
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The Distinguished Scientist, Burton Medal, OIA-MSA Outstanding Young Investigator and Outstanding Technologist Awards Nominations should include: 1) a letters from the primary MSA nominator describing the research accomplishments of the candidate with particular emphasis on the unique technical achievements in the Physical or Biological Sciences; and 2) supplemental letters of support from other members of the scientific community.
The Morton D. Maser Distinguished Service Award Nomination should include: 1) a letters from the primary MSA nominator describing the basis for the nomination; and 2) supplemental letters of support from other members of MSA.
The Deadline for receipt of Awards Nomination Packages is December 30, 1999. Please contact the MSA Business Office or Gracie Burke (mgburke-at-pitt.edu) for additional information.
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I think Warren Straszheim is right about an export program. If you have the Unix based system, which the RAS files suggests, I think you have access to an program called PBMPLUS. When I used the Unix IMIX, this was supplied in addition to the Imix software. The PGT documentation said this was a public domain program, by Jeff Poskanzer, which can convert RAS to several formats including TIFF. Look for PBMPLUS on your drives or ask PGT. good luck,
Dave Audette david.audette-at-sylvania.com
} -----Original Message----- } From: Dusevich, Vladimir [SMTP:DusevichV-at-umkc.edu] } Sent: Monday, October 18, 1999 11:34 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Imix PC pictures } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } After installation and working a lot with my new EDS systems (Imix PC) } from PGT, I am now preparing publications. And - surprise, surprise, - } I have found out that I cannot print good quality pictures (on } high quality printer which is not connected to our network)! } } The file format of stored images, RAS with overlays, cannot be read by } other programs. Of course, some of them can read RAS, but overlays } (with the most important information) are lost. } } The only advice I've got from PGT was to capture images from the monitor. } But then I will have files with much lower resolution than initial ones, } and this is certainly bad for high quality printing. } } All (if any) advises will be highly appreciated!!! } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 }
Email: bharesh_mandalia-at-madscientist.co.uk Name: Bharesh Mandalia Question: I am currently looking into blades which are coated with diamond like carbon (DLC). Thickness of this coating is about 2000A, on steel substrate. I examine the edge of these blades in a JEOL 6330F FEG SEM, at a tilt angle of 60deg. The magnification I look at is x20000. The edge appears round (ie large tip radius), but I increase the tilt to 80deg, the edge appears very sharp (ie very small tip radius (about 150A)).
What I'd Like to know what is happening as I increase the tilt from 60 to 80deg?
We have had excellent results using an Optronics DEI-750 CCD camera with a FlashPoint 128 video frame grabber for both fluorescence and bright field images. I understand the newer CCDs are even better and less expensive. The Optronics cameras are usually distributed via your Nikon and/or Olympus rep.
Best wishes,
Robyn Rufner, Ph.D. Director, The Center for Microscopy and Image Analysis Ross Hall, Suite 406 Adjunct Associate Professor, Anatomy and Cell Biology THE GEORGE WASHINGTON UNIVERSITY 2300 Eye Street, N.W., 431 Ross Hall Washington, DC 20037-2337 Voice: (202) 994-2881 Fax: (202) 994-8885 Internet: anarrr-at-gwumc.edu
This is interesting about metals blocking solidification of the JB-4... I knew this was true with osmium-fixed tissue, but until now, I didn't know exactly why tissue stained with alcian blue (which contains copper) didn't set up well. Just a comment though, the alcian blue stained tissue did eventually set up well enough to be sectioned months after it was embedded. Why? I have no idea.
On Mon, 18 Oct 1999 rmoretz-at-rdg.boehringer-ingelheim.com-at-Sparc5.Microscopy.Com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Aaron: } The methacrylate media generally do not work well in the presence of metals. } This is particularly true of osmium, and in my experience has included the } silver and gold-toned stains as well. Certainly your comment about the bulk } plastic polymerizing while the plastic immediately surrounding the stained } neurons not polymerizing would seem to corroborate my earlier anecdotal } experience. I too think that a soft epoxy (araldite is an excellent choice) } would be a preferable alternative. } } Roger Moretz } Dept of Toxicology } } } -----Original Message----- } } From: Aaron R. Best [SMTP:a_best-at-ou.edu] } } Sent: Friday, October 15, 1999 11:26 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: LM-Embedding Golgi-Cox in JB4 } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } We have not been able to embed Golg-Cox brain tissue in JB4. The JB4 } } solidifies, but the tissue within it does not. We have done it with and } } without polyethylene glycol. We have infiltrated the tissue with A+C } } for varying lengths of time. We have tried dehydrating the tissue in } } 100% ethanol prior to infiltration. We just purchased new JB4, so the } } solutions are not old. Any suggestions would be greatly appreciated. } }
Could you please help me to get some information on optical properties of cell membranes. I am a physicist and I met some problems during attempt to calibrate our equipment (photothermal microscope) for measuring living WBC properties. Could you advice some sources of information about:
1. What is absorption spectrum for intact WBC membranes in visible range? 2. Are there any natural photoactivated proteins? 3. What are light absorption wavelengths for them, range of interest is 400-600 nm? 4. What are molecular mechanisms for such photoactivation?
With thanks in advance
Dmitri Lapotko, Ph.D.
Luikov Heat and Mass Transfer Institute 15, Brovka Street Minsk, 220072 Belarus
We have been using P2O5 for over ten years and I think we're on our second bottle so the amount going into the environment is negligible (on our part). We have used silica and calcium dehydrants and they did not work as well or last as long. To increase the longevity of the dehydrant and aid in keeping the film and container dry we also used dry nitrogen gas to evacuate the desiccator, which is also connected to the scope for camera chamber evacuation. On a identical scope and desiccator with out the nitrogen we have much slower cycle times, and the desiccant is exhausted more quickly. The P2O5 is nasty to work with though so I will be interested in seeing what other chemicals people come up with.
I believe that the included responses to not address the problem. Which is that PGT (Princeton Gamma Tech) uses a special derivative of the Sun-RAS image file-format.
It is extremely easy to read the 16- or 8-bit version they use on a PC or Mac, if you are willing to neglect the overlayed micron-markers, text, etc.... which the user has included with PGT-specific software. Basically, you tell Photoshop, ImageTool, etc.... that the file is binary/raw, then you tell it the bit-depth, width, height, header-size (offset-to-data), big/little endian, black (high/low), etc....
The other technique to to print each image to "screen.ras", which will freeze the overlays on the image and create an 8-bit RAS-image. This file is easily read by most programs, since it in now in the generic Sun-RAS format. Also, PGT provides a convert-utility which will change this to PCX, JEOL-TIFF, TIFF, or JPEG.
Finally, please note that PGT will provide a script to dump multiple 16-bit RAS files to screen.ras, and then prepend the filename with "C_". If you use this script or if you manually dump to screen.ras, then it is extremely important that you only have "ONE" image display open. Otherwise, you will reduce the colour-depth of the "dumped" file.
I hope this helps!
regards,
Jim
} } } Sir, } } our software, analySIS, does read RAS images (SUN Raster Images). It can } then save them in a large number of other formats. If the number of } images is not too large, I could try to convert them for you. I can't } guarantee that it works, but maybe worth a shot. Let me know if you are } interested. Since we are not in the business of converting files, we are } not going to charge you for the conversion, but we will limit the time } we spend on this. } } Note to everybody: I am just trying to help Dr. Dusevich with his } images. Please do not send me any images for conversion without } discussing that with me first. I will not be responsible for images sent } to me without prior discussion. } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } } } } } Dear Volodya, } } Try InfanView (mailto:e9227474-at-student.tuwien.ac.at - I did not try this } } address). It is pretty good viewer and it may open RAS files. I did not try } } InfanView to print out, but you may try to save your image in some suitable } } format like "bmp" or "tiff" and print later from Photoshop for instance. } } Actually, I am really happy with that "viewer". It is very fast, recognizes } } most popular image formats, "slide show" option, direct viewing the } } Directory content etc. After a hard search of the Internet, I choose this } } program as a "default" viewer. InfanView is freeware, so, I have no any } } financial as well as other interest in this product. } } Good luck, Sergey. } } } } } } } } } } } } } After installation and working a lot with my new EDS systems (Imix PC) } } } from PGT, I am now preparing publications. And - surprise, surprise, - } } } I have found out that I cannot print good quality pictures (on } } } high quality printer which is not connected to our network)! } } } } } } The file format of stored images, RAS with overlays, cannot be read by } } } other programs. Of course, some of them can read RAS, but overlays } } } (with the most important information) are lost. } } } } } } The only advice I've got from PGT was to capture images from the } } } monitor. } } } But then I will have files with much lower resolution than initial ones, } } } and this is certainly bad for high quality printing. } } } } } } All (if any) advises will be highly appreciated!!! } } } } } } Vladimir M. Dusevich, Ph.D. } } } Electron Microscope Lab Manager } } } 3127 School of Dentistry } } } 650 E. 25th Street } } } Kansas City, MO 64108-2784 } } } } } } Phone: (816) 235-2072 } } } Fax: (816) 235-5524 } } }
Apart from its efficiency as a desiccant, one of the positive advantages of P2O5 as a desiccant for use in a vacuum system is that on exposure to moist air a "skin" rapidly forms on the surface of the powder, holding it together. This minimizes its redistribution in your vacuum system when the rough pumping. Some alternatives remain friable even when partially hydrated, and can scatter about. Compared with many other reagents used in electron microscopy P2O5 is fairly innocuous. Skin contamination is obviously not advisable, nor should the powder be breathed in, but the salts of P2O5 are constituents of NPK fertilizers and if you are worried about eutrophication of the local lake you could use the waste to fertilize your tomatos.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
} our software, analySIS, does read RAS images (SUN Raster Images). I
Unfortunately, that won't help in this case. The PGT image file format contains the annotation information in a proprietary trailer appended to the standard Sun raster format, so you'll just get the image without annotation, as would any other package which reads Sun image files.
The annotated image is generated internally as an encapsulated PostScript file before being sent to the printer. It may be possible to intercept the temp file and send that to an arbitrary network printer. Also, I am aware that writing the annotated image as a TIFF file is planned, but I don't know the release schedule.
No expert but I have been doing some of this lately. The first time I tried with osmicated tobacco seeds into LR White or unosmicated into LR Gold, it was so bad that it was funny. Despite a 3-4 day infiltration, the seeds literally popped out of the block face as if they were little bb's. In my next batch, I cut them into itty bitty wedges using a razor blade after the aldehyde fix stage. I then infiltrated slowly over 7 days. Minor improvement but I got some sections. Both plastics cut nicely at 0.5 um and thin sections look good initially but are so fragile. Sections that are initally 12 x 12 grid holes on a 400 mesh grid will usually burst over most grid holes and I end up with 1-4 grid holes surviving (immuno protocol but it happens early on so I think UA and Pb would do the same on their own). But they are pretty when they survive. Tom
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
It is difficult to understand what is your tilt angle means and why do you need to tilt, but I'd try a few things: - check for charging. If your blades a really thin, then coating could be too thick, but nevertheless it's worth trying. Even better to use environmental SEM with good resolution (not with BSE detector!). - use as low voltage as possible, and, at least not higher than 5 kV. Even better to use low voltage mode (I am not sure, but for your case maybe 0.6-0.8 kV will be OK) without any coating. - If nothing else works, try to break blades at LN temperature (of course, if steel is not austenitic) and take a look on profiles. - Look at blades "as is", before coating with DLC.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Phone: (816) 235-2072 Fax: (816) 235-5524
} -----Original Message----- } From: bharesh_mandalia-at-madscientist.co.uk } [mailto:bharesh_mandalia-at-madscientist.co.uk] } Sent: Tuesday, October 19, 1999 7:40 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: SEM Imaging Question: } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } Email: bharesh_mandalia-at-madscientist.co.uk } Name: Bharesh Mandalia } Question: I am currently looking into blades which are coated } with diamond } like carbon (DLC). Thickness of this coating is about 2000A, on steel } substrate. I examine the edge of these blades in a JEOL 6330F } FEG SEM, at a } tilt angle of 60deg. The magnification I look at is x20000. The edge } appears round (ie large tip radius), but I increase the tilt } to 80deg, the } edge appears very sharp (ie very small tip radius (about 150A)). } } What I'd Like to know what is happening as I increase the } tilt from 60 to } 80deg? } } Many thanks, } } Bharesh } } -------------------------------------------------------------- } ------------- } } }
PGT can supply a program to do a "screen" conversion on images so that they include any annotations. Then within the Applications Programs, the is a file conversion program which allows conversion from .ras to several other formats (including tif). You may then have to download the converted image to a floppy disk, but in the end you will have what you want. Additionally, you may want to look at some of your other Applications Programs, because you may be allele to save images in .tif to begin with if you do have the "PC" version of IMIX. Chris
We recently got some problems with our Gatan slow scan CCD camera in acquiring electron diffraction pattern, though everything looked normal when acquiring image. We tried varying the exposure time from 0.1 to 1 second, but there was nothing in the acquired image except the background similar to what appeared during gain reference acquisition. We even tried inserting the beam stop, but it did not appear in the pattern. Recently, one of our users changed the setting of image pattern type and image type. Does this have something to do with the anomaly in diffraction pattern acquisition?
Look forward to your help.
Regards,
Li Kun
Kun Li, Ph. D Institute of Materials Research and Engineering 3 Research Link Singapore 117603
I'm trying to help a student who is writting a PhD proposal on soil organic matter decomposition. As part of the study she would like to do some SEM studies in the first layers of the soil of a sugarcane plantation where no burning is applied before harvest. Is anyone doing anything like this? Can anyone recommend some literature or suggestions?
Thank you very much in advance
Adriana Rodriguez
******************************************************************* Adriana Pinheiro Martinelli Rodriguez Laboratorio de Biotecnologia Vegetal Av. Centenario 303, Cx. Postal 96 CENA/Universidade de Sao Paulo 13400-970, Piracicaba, SP, Brasil phone: +55-19- 429-4694 fax: +55-19- 429-4610 adriana-at-cena.usp.br http://www.cena.usp.br/labs/labbiotecveg.htm *******************************************************************
Bharesh: You can reduce this artifact by lowering the kV to the lowest which still gives reasonable resolution at the power required. Its an interesting phenomenon and a bit difficult to explain without a drawing. I'll try, just get a bit of paper and draw: 1 A vertical line, to symbolize the electron beam. (That vertical is 90 degrees in relation to the "normal" horizontal specimen position) 2 A sharp angle, of say 30 degrees, with the apex at the top and the vertical line entering at the apex. Draw the right side of angle at 60 degrees. The left side of the angle will happen to co-incide with the vertical.
3 Repeat that drawing, but draw the right side of the angle at 80 degrees. Now the left side will be 20 degrees off the vertical.
Electron emitted from the specimen come mostly from a hemispherical to pear shaped "penetration envelope", the region were most of the secondary electrons are formed. These envelopes are modeled and known as Monte Carlo patterns. Add to both sketches a drop shape with the pointed end at the electron beam entrance point. Ensure that the drop is vertical and symmetrical and projects at least on one side past the angle (which is your specimen).
There is your answer: The closer the penetrating envelope is to an open surface, the more electrons will be emitted from that spot and "sucked" onto the secondary detector. That is why a horizontal specimen is darker than one at a steep angle, that is why thin specimens (hairs) "glow" and that is why your specimen is apparently wider at the near vertical side. In all those cases the effect can be reduced by lowering kV or by using higher atomic number specimens; consider that a coating averages the atomic number of coating and actual specimen atomic number. Your "near" diamond coating for this exercise is just like amorphous carbon. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, October 19, 1999 10:40 PM, bharesh_mandalia-at-madscientist.co.uk [SMTP:bharesh_mandalia-at-madscientist.co.uk] wrote: } } } } Name: Bharesh Mandalia } Question: I am currently looking into blades which are coated with diamond } like carbon (DLC). Thickness of this coating is about 2000A, on steel } substrate. I examine the edge of these blades in a JEOL 6330F FEG SEM, at a } tilt angle of 60deg. The magnification I look at is x20000. The edge } appears round (ie large tip radius), but I increase the tilt to 80deg, the } edge appears very sharp (ie very small tip radius (about 150A)). } } What I'd Like to know what is happening as I increase the tilt from 60 to } 80deg? } } Many thanks, } } Bharesh } } --------------------------------------------------------------------------- } }
Hello, First of all many thanks to all who replied you made understanding a few quircks easier. I then asked what were the pros and cons of SEM with EDX+WDX vs E-probe.
1. You need a precise sample-spectrometer distance to get good analytical results and since SEM spectro are horizontally mounted and their focussing is imprecise (no optics) the quality of analyses suffers.
2. Tilt on SEM stage being not highly reproducible it will affect take- off angles of X-rays and hence affect ZAF corrections
3. Secondary electron detector in EPMA are disadvantaged because of their construction or, more often, their location limiting resolution and depth of field. New probes seems better in that respect.
4. EPMA because of the lack of tilting stage limits SEM observations to polished surfaces.
5. You can automate overnight analysis with an EPMA. SEM because of sequential WDS determinations are slower.
7. Personnal appreciations : a) Price of basic probe vs SEM equipped for analytical work is about 30% higher b) A respondant had SEM with EDX-WDX spectro (including a beam current meter) and claim good analytical results for their SEM mainly used when requiring a few elements in a few points. They also have a probe to compare. c) Another user claim that all he can deliver accurately with a WDS equipped SEM is qualitative analysis d) A user complained that the EDX of a SEM was difficult to reconcile with the WDS spectro because the difference in current requirement. e) Drift of several percent over an hour was observed on SEM needing frequent calibration and current monitoring. f) Probe being more complex you a dedicated operator to master the beast which will make or break the lab.
Hi, One of our divisions would like a simple and cheap, or free, ability to annotate and fix a micron bar on electronic images from a LM. They would like positionable alphanumeric labeling and a readable magnification indicator. Any ideas would be welcome. Thanks in advance, Russ Gillmeister, Xerox
Two questions: 1. Which vendor sells formvar + carbon coated grids which are also glow-discharged? 2. I know there are some methods which we can use to make the cytoskeleton more easily visible by TEM. Do any of you have the references at hand?
We have seen your message on the server. Please note we here at Electron Microscopy Sciences in Fort Washington Pa 215-646-1566 manufacture and make coated grids which are pre Glow Discharged. Please let us know if we can be of service to you. Sincerely, EMS 215-646-1566
This is in two parts. I've posted these recipes before. The first describes how to make scale markers like the transfer rub-ons. The second tells how to make scale markers for your optical microscope and have them ready.
Part 1 / 2 This is the recipe that I use in Photoshop 4.0 to put Black on White scale markers, text, and symbols onto micrographs. The results look just like the
rub-on transfers that I used to use.
1. create a layer (Photoshop 4.0 does this automatically with text tool) You must use Photoshop image mode for the layer option.
2. in that layer in the font and font size that you want, type the text. add a black line at an appropriate length and width and any other text, symbols, arrows, etc. that you want to put on the micrograph. By using the layer, you preserve the original micrograph in the background layer. you can use the info window to draw lines to particular lengths. If other layers are created when new text is added, merge those layers. Don't merge them with the background layer!
3. Select all (ctrl-A in the PC) The marquee will be around the whole layer.
4. You have to move the selected region up then down with an arrow key. (This is done in the PC with the Ctrl-shift-arrow key in the PC) what this
does is to select all of the objects in the layer individually. A Marquee should be around each object.
5. Select the foreground color as white. (You are going to write a white border around each Marquee.)
6. Go to Edit-Stroke and select the width of the white line you want (Width)
and select the Outside option. for 300 dpi images at about 4" x 5", I suggest a font size of about 14 (Arial) with a Width of about 3-4 pixels for
the stroke width. This will write a white border 4 pixels wide around all of the selected black features.
7. Deselect (ctrl-D)
8. If you want to save this as image in another format such as TIF or BMP, then you have to Merge the layers and save the image in that mode.
Note: you should have anti-aliasing selected for all this.
Part 2 / 2 Using Pre-drawn scale markers at different mags.
We use TWAIN import feature in Photoshop to bring images from two cameras on two stereomicroscopes. We have John Russ' Image Processing Toolkit for Photoshop. There is one plug-in, "IP*measure-Calibrate" that can be used to calibrate the image if a feature of known dimension is captured. It isn't absolutely needed to have this toolkit, but it made it a little easier. What I did was to take a digital image from a good metric ruler at each magnification setting of the microscope. (For an optical microscope with magnifications higher than those of a stereomicroscope, you will have to use another length standard.) I then calibrated the image, drew a scale marker on a new layer and labeled it for each setting. I changed the name of each layer to be indicative of the setting on the microscope that changed the mag. The only thing on each of these layers is the scale marker and the label. Once I had a layer for each of the different settings, I gathered them all into one photoshop file and labeled it with the appropriate microscope name. When I want to capture images from the microscope, I open Photoshop by opening this file of calibrated layers. After I collect the new image, I drag the appropriate layer from the open "calibrated scale markers" file onto the new image and align the scale marker where I want it.
A little work up front has saved me a lot of aggravation when it comes to calibrating images and putting scale markers on images. I plan to do the same thing for the mag settings on my TEM when I digitize them with my negative scanner, but I haven't invested the time yet.
-Scott
-----Original Message----- } From: Gillmeister, Russ [mailto:RGillmeister-at-sdms.usa.xerox.com] Sent: Wednesday, October 20, 1999 10:51 AM To: 'MSA'
Hi, One of our divisions would like a simple and cheap, or free, ability to annotate and fix a micron bar on electronic images from a LM. They would like positionable alphanumeric labeling and a readable magnification indicator. Any ideas would be welcome. Thanks in advance, Russ Gillmeister, Xerox
Thanks for the summary. All the points made are very true when considering probe vs. SEM/WDS. I was going to reply ealier, but forgot. Here are my two cents.
We purchased a WDS system a few years ago because we could not justify the high price tag for a probe for the limited quantitative needs. Luckily our SEM had a WDS button which knocks out one of the condenser lenses for higher beam currents. Horizontrally mounted WDS are much less sensitive to sampling height position relative to vertically mounted ones, but nontheless, it must remain the same during the analysis. We had our service engineering hook up leads from the final focussing (objective) lens so that we can measure the voltage on the lens as a function of working distance. Removing a port on the specimen chamber, we physically set our wd and then without moving the stage, set our operating conditions for both EDS and WDS over a range of operating voltages/beam currents. Once established we reorded the obj voltage for each.
As for the SEM stage, again we were lucky that the manufactured stage had a mount that they hung the tilting and rotation axis on. We removed this inner stage and axis connections and had our shop make a new holder for mounting flat plates to. We had a series of plates made to hold about every type of sample we used, polished epoxy mounts, geological thin sections, and of course our standards.
The beam current is measured from a retractable homemade faraday cup. Since our wd is 40 mm, it is easily inserted above the sample while the sample is in the analytical position. Yes it is a pain to do it manually, but...... We have found that under proper gun conditions the beam drift is less than 2% at 30 nA over an eight hour time period. The other problem with beam current stability that wasn't addresed in your responses is the LaB6 vs. W guns. It is well known that W is a more stable source, so one needs to change emitters depending on imaging resolution/qualitative microanalysis and quantitative analysis.
One mistake we made was to think we could do EDS and WDS simutaniously at WDS beam currents. Wrong. Even when we put an aperture in front of the EDS detector, the EDS background was swamped with incoming bse's, producing a hump at a much higher energy range than found with standard Be window detectors. So it's only good for quick look at major peaks in EDS under these conditions.
Hope this helps. Lou Ross Senior Electron Microscope Specialist Room 101 Department of Geological Sciences University of Missouri Columbia, MO 65211 (573) 882-4777, fax 882=5458 email: rosslm-at-missouri.edu web: www.missouri.edu/~geosclmr/ebaf.html
I cannot address the problem with your printer. We have a photographic quality printer and it prints PGT *.ras images fine. You might want to try printing test images to verify the printer is working properly.
With the IMIX, you can save the images as *.tif files using the File "Save As" option. The annotation will not be saved. Add annotation with other software, such as,. Word or Photoshop. It is a little inconvenient, but while in Photoshop you can adjust your Gamma.
Note, Optimas (Media Cybernetics) will read *.ras image files, annotation is not read with the image.
John Catino Minerals Technologies
"Dusevich, Vladimir" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } After installation and working a lot with my new EDS systems (Imix PC) } from PGT, I am now preparing publications. And - surprise, surprise, - } I have found out that I cannot print good quality pictures (on } high quality printer which is not connected to our network)! } } The file format of stored images, RAS with overlays, cannot be read by } other programs. Of course, some of them can read RAS, but overlays } (with the most important information) are lost. } } The only advice I've got from PGT was to capture images from the monitor. } But then I will have files with much lower resolution than initial ones, } and this is certainly bad for high quality printing. } } All (if any) advises will be highly appreciated!!! } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 }
Yuhui: 1 Butvar filmed (or substrates) grids are stable under the electron beam. They don't need carbon and so they do not need glow discharge treatment.
2 From memory: Cytoskeletons are destroyed by cold fixation. I am sure there are other things to that and I don't remember the reference.
Disclaimer: ProSciTech supplies Butvar coated grids. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, October 21, 1999 2:07 AM, Yuhui_Xu [SMTP:Yuhui_Xu-at-hms.harvard.edu] wrote: } } } Dear collegues: } } Two questions: } 1. Which vendor sells formvar + carbon coated grids which are also } glow-discharged? } 2. I know there are some methods which we can use to make the cytoskeleton } more easily visible by TEM. Do any of you have the references at hand? } } Thanks for a reply! } Regards, } Yuhui } } } Yuhui Xu, MD,PhD }
Yuhui Xu asked: ================================================ Two questions: 1. Which vendor sells formvar + carbon coated grids which are also glow-discharged? ================================================= SPI Supplies, since 1975, has supplied to TEM users Formvar® as well as carbon coated (filmed) grids and when the carbon needs to be more hydrophilic, the grids can be "glow discharge" treated. However, this effect is not permanent, and the treated grids should be used within about two weeks of treatment (more or less) since the effect does start to wear off.
Full details on this service can be found on our website given below.
Chuck
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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.spi.cc ######################## ============================================
I'm looking for advice from anybody that has successfully fixed fungal yeast cells in animal tissue. I am currently working with a thick-walled yeast that tends to crush in the tissue because of reasons unknown. I have played with the osmolarity of the fixative, as well as the dehydration time, both to no avail. The fixitives I have tried are trump's(4F:1G), and a 2.5% glut. fixitive. Some of the cells improved with the addition of sucrose to the fixitive, but there was still alot of crushed, hollow cells. If anybody has advice on the process or a good recipe for this kind of sample it would be appreciated.
Eric Tarcha Medical Technology Michigan State University tarchaer-at-msu.edu
Several of you listers expressed interest in the new "toy" microscope when it was announced earlier this year. I made several attempts to get more information from both Mattel & Intel, with no success. It's now on the market, and a review appeared in the N.Y. Times last week; you can read it at http://www.nytimes.com/library/tech/99/10/circuits/articles/14pets.html It works on a Windows computer with a USB port, and I'm a Mac person, so I'm hoping that if any of you get one that you'll share your impressions with the rest of us.
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I have never had to embed tissue in Spurr, since I do SEM, however I have a kit since I use it to make replicas for SEM. Now, one of our faculty has asked me to embed some tissue samples for him today. They are currently in Formalin. Does anyone have a (hopefully) quick and easy protocol for this?
} I think I saw this on a computer show. If you search the Net at ZDTV and } check their products tested section it should show it. It seemed pretty } neat for a toy. Cost was ~$100. Hope this helps, Steve.
Steve
You've got the price right. The scope is a cheap plastic one, but I'm wondering 1) if the camera can be removed to use on something better, and 2) if it's as good as Snappy/QuickCam cameras.
Caroline
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
For how long are you fixing? Have you tried extending the time in primary and/or secondary fixative? Maybe the yeast just aren't getting fixed well enough?
Tamara Howard CSHL
On Thu, 21 Oct 1999, Eric J Tarcha wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm looking for advice from anybody that has successfully fixed fungal yeast } cells in animal tissue. I am currently working with a thick-walled yeast that } tends to crush in the tissue because of reasons unknown. I have played with } the osmolarity of the fixative, as well as the dehydration time, both to no } avail. The fixitives I have tried are trump's(4F:1G), and a 2.5% glut. } fixitive. Some of the cells improved with the addition of sucrose to the } fixitive, but there was still alot of crushed, hollow cells. If anybody has } advice on the process or a good recipe for this kind of sample it would be } appreciated. } } Eric Tarcha } Medical Technology } Michigan State University } tarchaer-at-msu.edu } } }
"Cargille Optical Liquids and Mounting Media for the Microscopist "
Speaker is Robert Sacher of Cargille Laboratories.
Robert Sacher will speak about Cargille Refractive Index Liquids and the different techniques for using them. He will discuss the Cargille Microscope Immersion Oils and there fluorescence and viscosities. Plus he will discuss the Cargille Meltmount Mounting Media and their different refractive indices. Issue of toxicity will be considered as well as the significance of the Becke line, etc. This is a great opportunity to learn basic information about optical liquids and mounting media; understanding how to use these properly is a key to obtaining the best possible image through a light microscope. There will be ample time for questions and answers
Dear Listers, } I have a customer studying nerves at the TEM level in crayfish. He has given me the } antenna to cut and because of the thick cuticle, good fixation & infiltration is a real problem. The tissue was infiltrated over a 3 day period with increasing amounts of resin and embedded in an embed 812/Araldite mixture. The embedded sample was holey and the tissue that did remain had terrible morphology. Does anyone out there have any suggestions as to how to improve morphology and get the tissue well infiltrated? } Any ideas will really be appreciated. } Thank you, } Mary Gail Engle } Electron Microscopy & Imaging Facility } University of Kentucky }
We are looking into replacing an old Galai CUE-4 system. The new set up should include a new CCD camera attached to our light microscope, "feeding" single frames to a Macintosh, possibly one of the new iMacs or a G3/G4 computer. The Mac platform is the one of choice, as we use Macs to run DigitalMicrograph with our TEM CCD camera, and to process images.
I wonder whether any of you have a set-up similar to what we would like to install in our lab, and could give us some advice.
Thanks!
Ishi Talmon Dept. Chem. Engng. Technion-Israel Inst. of Technology
MicroMaterials, Europe's technology leader in Nano / Micro-mechanical Properties has changed distributors in the North America to Molecular Imaging.
At AVS / Seattle next week Dr. Jim Smith, Founder of MicroMaterials will be hold technical talks on application of state of the art in mechanical properties testing.
I invite you to join us at Molecular Imaging's Booth # 938 & 940.
RSVP
George Sibbald, CEO Molecular Imaging www.molec.com
} You've got the price right. The scope is a cheap plastic one, but I'm } wondering 1) if the camera can be removed to use on something better, and } 2) if it's as good as Snappy/QuickCam cameras.
The NYTimes review indicated that the camera was removeable for "macro" work but not if it could be fit to another scope. The review also failed to mention anything about resolution except that it seemed, subjectively, pretty good to the unsophisiticated reviewer.
Yuhui Xu asked: =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D Two questions: 1. Which vendor sells formvar + carbon coated grids which are also glow-discharged =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D Hi Yuhui, We make our own grids, means carbon coat them & also glow them before use.We had the same problem of grids getting hydrophobic after 2 weeks. Try this it works,after you glow them store grids in the refrigerator till you need it .If you do this the grids remain hydrophilic even after a month.Get your grids out before you need them & when your are done stick them back in the refrigerator.
Reena Zalpuri EM Lab UC Berkeley E-Mail jubu-at-uclink4.berkeley.edu
Anybody out there got a SM-PCD40 Probe Current Detector attachment for an 840 that they want to sell, or maybe want to trade for a JEOL/Varian LaB6 setup for 840?
thanks
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
} I'm looking for advice from anybody that has successfully fixed fungal } yeast } cells in animal tissue. I am currently working with a thick-walled } yeast that } tends to crush in the tissue because of reasons unknown. I have } played with } the osmolarity of the fixative, as well as the dehydration time, both } to no } avail. The fixitives I have tried are trump's(4F:1G), and a 2.5% } glut. } fixitive. Some of the cells improved with the addition of sucrose to } the } fixitive, but there was still alot of crushed, hollow cells. If } anybody has } advice on the process or a good recipe for this kind of sample it } would be } appreciated. } } Eric Tarcha } Medical Technology } Michigan State University } tarchaer-at-msu.edu
I know nothing about fungi in animal tissue, but a lot more about fungi in plant tissue. Fungal cell walls seem to be less permeable to fixatives than plant cell walls and , of course, no comparison to animal cells, also, young cells are easier to prepare than older stages. Are you sure that the yeast cells are alive and in good condition in the animal tissue?
Our prepartion protocoll for fungal infected plant tissue is: 2,5% buffered glutaraldehyde for 1 h, washing steps, then 0.5 -1% Osmiumteroxide, buffered, for 1 h at room temperature. Dehydration in ethanol or acetone, embedding in LR-White or Epon. We get better structural preservation if we dehydrate in the cold (PLT-Method): 1,5 h at zero (Grad Celcius) in 30% Ethanol, 1,5 at -20 in 50% Ethanol, overnight at -35 in 70% EtOH, 1,5 h at -35 in 100% EtOH. We cut very small samples if the tissue is difficult. Good Luck!
Anne Heller
-- Dr. Anne Heller, AG Elektronenmikroskopie, Institut f=FCr Botanik (210), Universit=E4t Hohenheim, Garbenstra=DFe 30 D-70593 Stuttgart heller-at-uni-hohenheim.de
Dear all, once again I put this question on the board, in hope that I get more answers as before the crash of nestors pc at the end of september. We search for a protocol or tips and experiences in histochemical localization of proteases on semi- or ultrathin sections. We search for substances, which could act as substrates for proteases and would make it possible to visualize the endproduct at the localization of the enzymatic process (for example precipitation). Best regards, Bernward Laube
Applications are being accepted (deadline Jan 2000) for NRC Post Doc slots at NIST to begin Oct. 2000 for 2 years. See the web page for details http://www.nist.gov/oiaa/pdfront.htm
Gary, since accuracy of analyses is paramount, you'll need to give consideration to beam stability. Because normal LaB6 cathodes emit from a very small microflat at the apex of the crystal's cone, beam stability will be less than a filament's. I should quickly add that for visual and photomicrographic purposes beam stability, even in TEM is not an issue or a problem.
However, during typical analyses times of 120 seconds the picoamp reading can drift with the smallest cathodes flats of 15 or 20 square micrometers. These happen to be the most purchased since smaller flats result in greater brightness. Unfortunately the larger 40 flat made by Kimball costs a couple hundred more US$. The larger flat is more stable and in an SEM would not affect resolution or compromise brightness. Here is just another little dilemma for you on the path to quantitative analyses. 1 Battle on with more frequent beam adjustments and filament centering 2 Pay up for a more suitable cathode with the larger flat. 3 Change Wehnelt units (got a spare?) for one with a filament when performing critical analyses 4 Sacrifice the beloved LaB6 and use filaments only. Lucky Gary, you have choices! Disclaimer: PST supplies Kimball LaB6 cathodes and filaments (not to N America). Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, October 21, 1999 6:00 AM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote:
} Hello listers: } } I am looking to add an X-ray EDS system to my LaB6 SEM. } Feedback from users would be appreciated. } } Here are my main details: } } No LN2 (cryo cooled) } Application is determination of composition of microcircuit areas: } 1) passivation (PSG, nitride ) } 2) metal (Al/Si) } 3)poly-Si (poly Silicon) } 4) dry oxide (SiO2) } } would like to run on a PC with Win95/98. Ease of use is important, } price is not a major factor. Sensitivity and accuracy are paramount. } } thanks, } gary g. }
First, I'd like to say that tissue fixed in formalin often doesn't look very good at TEM-levels.
My method is not quick, but here it is anyway incase no one has a quick way:
1. Put tissue (my tissue is 5mm X 2.5mm X 2.4mm in size) into saline or buffer after the formalin fix. The saline/buffer should be the same temp. as the fix, fast changes in temp. can cause buckling of membranes. Put samples on a rotator of some kind for 30 min.
2. Change the solution to 50% ethanol for 30 min. (leaving on the rotator to allow for faster infiltration)
3. Follow this by 4-15 min. changes in: 70%, 95%, and 100% ethanol respectively.
4. Place the tissue into propylene oxide for 20 min.
5. Place the tissue into propylene oxide/ Spurrs (1 to 1) for 2 hrs.
6. Place into 1 part propylene oxide/ 2 parts Spurss for 2 hrs. or longer (can go overnite)
7. Place into 100% Spurrs 4 hrs. or longer
8. Place into fresh Spurrs and embedd (I put them into a 45 degree C oven for 3 days, then into a 60 degree oven for one day).
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have never had to embed tissue in Spurr, since I do SEM, however I have } a kit since I use it to make replicas for SEM. Now, one of our faculty } has asked me to embed some tissue samples for him today. They are } currently in Formalin. Does anyone have a (hopefully) quick and easy } protocol for this? } } Thanks in advance. } } Ron } lherault-at-bu.edu } } } } } }
Electron -Microscopist Postdoc Fellowship: call for Applicants
A fellowship is available at the laboratory of Electronic Materials and Engineering (EME), University of Barcelona. A PhD or Similar Experience in the field of Electron Microscopy and related techniques is needed (preferable in the field of semiconductor materials and structures for devices).
The candidate will collaborate in TEM analysis of semiconductors and electronic ceramics. In principle, duration of the fellowship is one year, but it can be extended to a second year. Selected candidate ought to join the group on an immediate basis.
People interested is required to contact, before Novembre 15th to the following address:
Dr. Alejandro Perez-Rodriguez or Dr. Albert Romano-Rodriguez Departament d'Electronica Facultat de Fisica Universitat de Barcelona c/ Marti i Franques, 1 E-08028 Barcelona, SPAIN Tel 34 93 4029069/4021147 Fax 34 93 4021148 e-mail: perez-ro-at-el.ub.es, romano-at-el.ub.es Prof. Albert Romano-Rodriguez Dept. of Electronics Faculty of Physics University of Barcelona c/ Marti i Franques, 1 E-08028 BARCELONA Spain tel: +34-93-402 90 69 FAX: +34-93-402 11 48 e-mail: romano-at-el.ub.es
Eric, While I have extensive experience with yeast, I have little experience with fixing/embedding yeast within animal tissue. However,I would bet that your problem results from poor infiltration of your resin. The yeast cell wall is very difficult to get resin through and yeast cells that are processed with their walls on often fall out of the sections or appear 'crushed'. One thing to try is a low viscosity resin. Also, when we embed yeast with their walls on we treat them with sodium metaperiodate, this treatment breaks some of the wall linkages making it more permeable to the resin (LR White or Spurrs in our case). I don't know what metaperiodate treatment will do to your animal tissue - probably nothing. If you want to give it a try, you can check out the protocol at genome-www.stanford.edu/group/botlab Good luck
Jon Mulholland Botstein lab/ Stanford Univ.
On Thu, 21 Oct 1999, Eric J Tarcha wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm looking for advice from anybody that has successfully fixed fungal yeast } cells in animal tissue. I am currently working with a thick-walled yeast that } tends to crush in the tissue because of reasons unknown. I have played with } the osmolarity of the fixative, as well as the dehydration time, both to no } avail. The fixitives I have tried are trump's(4F:1G), and a 2.5% glut. } fixitive. Some of the cells improved with the addition of sucrose to the } fixitive, but there was still alot of crushed, hollow cells. If anybody has } advice on the process or a good recipe for this kind of sample it would be } appreciated. } } Eric Tarcha } Medical Technology } Michigan State University } tarchaer-at-msu.edu } }
} You could just abandon the dessicant and rely on the rotary pump....works } for us on a number of systems, and I'd be doubtful of how much extra } advantage you get from the dessicant, especially in a short time. } Sally Stowe }
The same is true for our EM. Since over 9 years, films are dried over night by means of a rotary pump, without using any desiccant. Works fine for us - even for cryo EM.
Dr. Reinhard Rachel Universitaet Regensburg Lehrstuhl fuer Mikrobiologie (Prof. Dr. K.O. Stetter) D - 93040 Regensburg Tel.: +49-941-943-4534 Fax.: +49-941-943-1824 e-mail: Reinhard.Rachel-at-biologie.uni-regensburg.de
Ron: My protocol to get to Spurr's is as follows: post fix in osmium 45 min 50% ethanol 10 min 70% eth 10 min 95% eth 10 min 2 changes 100% eth 10 min each 1:1 Spurr's/ 100% eth 10 min spurr's 15 min spurr's 10 min Cure overnight at 70 deg. C Don't know how "quick and easy" it wil be compared to others protocols, but I find it easy to use and the time fits nicely into my lab's routine. Hope it helps, Wanda Shotsberger Harris Methodist FW Fort Worth TX
---------- } From: Ron L'Herault To: 'Microscopy-at-sparc5.microscopy.com' -----------------------------------------------------------------------.
I have never had to embed tissue in Spurr, since I do SEM, however I have a kit since I use it to make replicas for SEM. Now, one of our faculty has asked me to embed some tissue samples for him today. They are currently in Formalin. Does anyone have a (hopefully) quick and easy protocol for this?
Hi, Does anyone have any advice on embedding media for immunogold labeling of proteins at the TEM level? I am preparing plant tissue for immunolabeling, and weighing my options carefully. I have used medium grade LR White polymerized at -20C with UV in the past, and found that although the antigenicity of the tissue is markedly superior to Spurr's embeded tissue, the quality of tissue preservation/appearance leaves a lot to be desired. I've recently come across two methods that give me pause. One uses soft grade LR White polymerized at 60C and shows micrographs of tissue that greatly resembles Spurr's or Epon/Araldite embedded tissue. Could the soft grade be the difference? The second (Brorson, S-H. Micron. 29(2/3):89) reports that high-accelerator epoxy resin plus antigen retrieval results in immunolabeling as good, if not better than LR White with preservation/appearance of tissues equal to epoxy. Has anyone tried this in general with plant tissue or tried applying to other resins (the author of that paper uses LX-112)? I would appreciate any advice that you have on the subject. Thanks, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
We have a system simular to what you have discribed. Photometrics Sensys CCD camera and capturing single or series images via a power mac with IPLab (Scanalytics software) or photoshop plugin. When we upgrade we would like to go to a G4 Mac and a fire wire camera. We have been extremely happy with the Photometrics/Mac combination. Very little down time.
Bob U of Washington Morphology Core Lab
On Thu, 21 Oct 1999, Prof. Ishi Talmon wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopists, } } We are looking into replacing an old Galai CUE-4 system. The new set up } should include a new CCD camera attached to our light microscope, } "feeding" single frames to a Macintosh, possibly one of the new iMacs or a } G3/G4 computer. The Mac platform is the one of choice, as we use Macs to } run DigitalMicrograph with our TEM CCD camera, and to process images. } } I wonder whether any of you have a set-up similar to what we would like to } install in our lab, and could give us some advice. } } Thanks! } } Ishi Talmon } Dept. Chem. Engng. } Technion-Israel Inst. of Technology } } } }
This method is fine for monolayers and very small slivers of tissue, but infiltration times need to be lengthened for larger pieces of tissue and anything with cell walls, such as yeasts. For 1 mm cubed biopsies, we dehydrate 2x in 100% EtOH for 15 min (more changes for yeasts/plants), and do 2 changes of 100% Spurr for at least 30 min before fresh Spurr and baking. Bigger tissue pieces, and hard-to embed materials, need more changes and longer times.
On Fri, 22 Oct 1999, Shotsberger-Gray, Wanda wrote:
} Date: Fri, 22 Oct 1999 10:07:00 -0500 } From: Shotsberger-Gray, Wanda {WandaShotsberger-Gray-at-hmhs.com} } To: Ron L'Herault {lherault-at-bu.edu} , } "'Microscopy-at-sparc5.microscopy.com'" {Microscopy-at-sparc5.microscopy.com} } Subject: RE: Spurr embedding } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ron: } My protocol to get to Spurr's is as follows: } post fix in osmium 45 min } 50% ethanol 10 min } 70% eth 10 min } 95% eth 10 min } 2 changes 100% eth 10 min each } 1:1 Spurr's/ 100% eth 10 min } spurr's 15 min } spurr's 10 min } Cure overnight at 70 deg. C } Don't know how "quick and easy" it wil be compared to others protocols, but } I find it easy to use and the time fits nicely into my lab's routine. } Hope it helps, } Wanda Shotsberger } Harris Methodist FW } Fort Worth TX } } ---------- } } From: Ron L'Herault } To: 'Microscopy-at-sparc5.microscopy.com' } Subject: Spurr embedding } Date: Thursday, October 21, 1999 9:24AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have never had to embed tissue in Spurr, since I do SEM, however I have } a kit since I use it to make replicas for SEM. Now, one of our faculty } has asked me to embed some tissue samples for him today. They are } currently in Formalin. Does anyone have a (hopefully) quick and easy } protocol for this? } } Thanks in advance. } } Ron } lherault-at-bu.edu } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Thanks to all who sent me Spurr protocols. I won't know how sucessful I was until later in the week. I'll keep the group posted.
Ron ----- Original Message ----- } From: Shotsberger-Gray, Wanda {WandaShotsberger-Gray-at-hmhs.com} To: Ron L'Herault {lherault-at-bu.edu} ; {Microscopy-at-sparc5.microscopy.com} Sent: Friday, October 22, 1999 11:07 AM
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There has been a huge rash of spam coming down the lines the last few days. I expect even more with the end of the year holidays coming since I've noticed alot of "new" bulk emailers sites are turning up. They are mainly trying to sell you "stuff".
Just for your own information, I keep a records/files of the spammer's Email domain names and KeyWords used by the spammers. These files are check by the filter each time a message comes through for the server. If a message comes from a potential host which has sent or relayed spam (and that particuliar address is not on the "exceptions list") or a suspect keyword is found then the message is rejected and an Email message sent back to the originator. The exceptions file is a list of addresses which for various reasons trigger the filter but are actually valid subscribers. If the filtering software sees that an address is on exceptions list then the message is allowed to pass through.
Since one does not know in advance all possible addresses of spammers and new ones are creeping in everyday, some spam will always get through. As I see a new "address" it is added to the file and the same site should not (in principle) get through a second time.
Occasionally a few of you inadvertantly caught because of a match of similiar domain names, key words in the subject lines, etc... If this happens please follow the directions in the return mail. I will see to it that your "individual" address is added to the exceptions so that your posts can get through. or advise your of what needs to be done.BTW it is not realistic to put the entire subscription list on the exceptions file since the subscribers list changes daily.
Also while I've got your attention please remember 2 other things.
1.) Never send an attachment of any kind. 2.) Never set your Email program to send embedded HTML this is for 2 reasons a). Text only Email programs get mounds of {html} code in the message which makes it almost unreadable. b.) Embedded html is sometimes sent with code that fingers the message has having an "attachment" this will be picked up by the filter and your message will be rejected Microsoft Outlook Express Users are the most likely to have this last problem/option . You may disable it under your references options just set your program to SEND in PLAIN TEXT not HTML.
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Mary Gail Try to get the youngest (smallest) crayfish possible and cut off the antenna just after the animal molts. Cut cross-sections of the antennae and place them in the fix on a stirrer-shaker. A stirrer-shaker for processing and then vacuum infiltration during embedding should be helpful. Joan
On Thu, 21 Oct 1999, Mary Gail Engle wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } I have a customer studying nerves at the TEM level in crayfish. He has } given me the } } antenna to cut and because of the thick cuticle, good fixation & } infiltration is a real problem. The tissue was infiltrated over a 3 day } period with increasing amounts of resin and embedded in an embed } 812/Araldite mixture. The embedded sample was holey and the tissue that did } remain had terrible morphology. Does anyone out there have any suggestions } as to how to improve morphology and get the tissue well infiltrated? } } Any ideas will really be appreciated. } } Thank you, } } Mary Gail Engle } } Electron Microscopy & Imaging Facility } } University of Kentucky } } } } }
Dear List, Is there any problem with fixing and embedding macrophages using standard protocols (GA,etoh,propylene oxide, P.O.parts epon etc), because of enzymes etc on the surface. We seem to have fix/infiltration problems.
Hi all, we have just discovered that the isolation valve on our In-lens field emission Hitachi Model S5000 has a leak. Hitachi engineers recommend that we replace the unit. Potential cost $20K(aus). The fault may simply be a piece of dust on the O-ring. I am, as you may imagine, fairly reluctant to order one and find I do not need it.
Has anyone else run in to a similar problem with their system? How easy is it to dismantle the S5000 column and check the isolation valve O-ring?
Thanks in advance. Colin MacRae ************************************************************************ Electron Microscopy Group
CSIRO Minerals Colin.MacRae-at-minerals.csiro.au
O Box 312, Clayton South, Ph. 61 3 9545 8800 Vic, 3169 Fax 61 3 9562 8919 AUSTRALIA
EM units WWW site http://www.minerals.csiro.au/em-unit/ *************************************************************************
It is a tedious but relatively easy procedure not to be undertaken by the inexperienced, incompetant and/or faint of heart . I just replaced one and the cost was 1/2 day of replacment and three days of pumping down. After explaining to Hitachi what I had done, they "shook their heads".
First, order the parts: "O" ring, two 6 inch conflats flanges, four 3/4 inch conflats. Use ultra-high vacuum techniques: clean tools and gloves in a clean environment, cover everything with aluminum foil when not re-assembling. The idea is to dis-assemble and remove the column in one piece below the isolation valve.
1. Turn off ion pumps. 2. Open all three side manifold valves.(right side of column) 3. Vent gun and specimen chamber. 4. Once fully vented, turn off diff pump or turbo pump. 5. Turn off "display power" and "evacuation power". Unplug SEM. 6. Remove High voltage cable and column shrouds. 7. Remove ion pump cables. 8. From SEM rear, remove ion pump magnets and heaters and related mounting hardware. 9. Dis-connect remaining heater wires. 10. Disconnect the vacuum manifold at the bottom valve. 11. Look for any remaining hardware that would prevent you from removing the column.
At this point we need to lift the column from the SEM. I use a 3 ft X 4 ft ( 50 cm X 90 cm ?) cart, covered it with foil and placed it next to the column. I usually clean all the column from dust using a dry paintbrush then follow with a towel immersed in alcohol.
12. Locate the four column nuts located below the isolation valve but above the specimen chamber. There is a "recess" at this area for these nuts. 13. Remove the two nuts closest to the rear of the SEM. (12 -13 mm open end wrench). 14. Using four people: three to lift, one to unbolt the remaining two nuts while supporting the rear of the column assembly. Do not let the column tilt unless necessary. 15. Lift column, ion pumps without magnets, etc onto the table. 16. Support the Column using whatever packing foam etc. Cover with foil to avoid dust. 17. Remove the sixteen nuts from the column bottom, exposing the isolation valve and replace the "O" ring. 18. Clean all isolation valve area.
Reverse above procedure and re-assemble column. Pump down and rebake the column in accordance with the procedure outlined in the Hitachi manual.
A word of caution in several areas: Do NOT attempt to access the isolation valve from the side. The isolation valve is NOT accessible here. You may damage the bellows. Do NOT adjust the isolation valve rod. It is factory adjusted and should only be adjusted if the rod is replaced. If it is adjusted it is not the end of the world just tedius to adjust. Use common sense: do not place column on it side so as to bend the isolation valve or otyher components.
I should probably put a disclaimer here that the above procedure is done at you own risk . You would be surprised at the people who ask for advice only to screw it up then expect you to fix it for free.But then again I live in California home of the free and attorneys.
Contact me if you need other info.
Good luck,
Earl
Colin MacRae wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all, } we have just discovered that the isolation valve on our In-lens field } emission Hitachi Model S5000 has a leak. Hitachi engineers recommend that } we replace the unit. Potential cost $20K(aus). The fault may simply be a } piece of dust on the O-ring. I am, as you may imagine, fairly reluctant to } order one and find I do not need it. } } Has anyone else run in to a similar problem with their system? How easy is } it to dismantle the S5000 column and check the isolation valve O-ring? } } Thanks in advance. } Colin MacRae } ************************************************************************ } Electron Microscopy Group } } CSIRO } Minerals Colin.MacRae-at-minerals.csiro.au } } O Box 312, Clayton South, Ph. 61 3 9545 8800 } Vic, 3169 Fax 61 3 9562 8919 } AUSTRALIA } } EM units WWW site http://www.minerals.csiro.au/em-unit/ } *************************************************************************
The University of Michigan Materials Science and Engineering Department has the following machine available.
Hitachi S-800 FEG SEM with GW Electronics Microchannel Plate BSE detector and Macintosh-based 4pi Analysis digital image acquisition system. Instrument purchased new in 1988 and maintained under service contract until Sept., 1999: $70,000. For further information call (734) 764-3357.
_______________________________________ Anne E. Huber Ph.D., Instrument Analyst Materials Science and Engineering Dept. The University of Michigan 2300 Hayward St. Ann Arbor, MI 48109-2136 ahuber-at-umich.edu (734)764-3357 _______________________________________
Can anyone recommend a diamond knife suitable for obtaining thin sections of Iron catalyst particles ? I currently have a Microstar SU knife with 2 mm length and 45 degree angle. After using it only once on the catalyst powder embedded in spurr's resin, I found that the edge was already scratched. Did I choose the wrong knife for this kind of work ?
Sincerely W. Erasmus
Willem Erasmus Snr. Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 9604211 Fax : +27 +16 9602826 E-mail : willem.erasmus-at-sasol.com
Hello All This is probably one of those questions where everyone has there own version but is 1% and 2% OsO4 in phosphate buffer solutions stable at cool (20-22 C) room temperature? We want to store the double container- para filmed solutions in the hood so we can do away with an old refrigerator but the the only other refrigerator for the use of buffers and glutaraldehyde chemicals is shared with an immuno-histotech that logically does not want the osmium in there at all. SPI alludes to keeping the diluted stock refrigerated, but the MSDS's we have states room temp or cooler. Our safety people don't know except what the MSDS says. Thanks ahead for any help. P.S. Does CAP regulations have any additional requirements?
Here is another source for a review of the Intel/Mattel QX3 Computer Microscope.
San Jose Mercury News, 10/24/99, page 1E, or www.siliconvalley.com/computing.
The article is by Mike Langberg, Merc's computing editor. It has a couple of pictures. Title is something like 'Kids' microscope doesn't bear close inspection'. He didn't like it, thought a conventional microscope gave kids a better view of the micro world. I thought he might have been kind of hard on it, but to each his/her own.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Listers, Could someone please point me towards a reference for measuring the thickness of a Carbon evaporative coat on polished brass. I have tried a search and found nothing. I am looking for something I can cite in a proceedure, and thought I saw one listed here some time ago.
Sarah A. W. Lundberg Microbeam Facility Analyst Office (702) 895-1134 University of Nevada, Las Vegas Lab (702) 895-2660 4505 Maryland Parkway Box 454010 email lundberg-at-nevada.edu Las Vegas, NV 89154-4010 Fax (702) 895-4064
Home : 4489 De Forest Street Las Vegas, NV 89103 (702) 871-9635
Sarah A. W. Lundberg Microbeam Facility Analyst Office (702) 895-1134 University of Nevada, Las Vegas Lab (702) 895-2660 4505 Maryland Parkway Box 454010 email lundberg-at-nevada.edu Las Vegas, NV 89154-4010 Fax (702) 895-4064
Home : 4489 De Forest Street Las Vegas, NV 89103 (702) 871-9635
Personally, I prefer to use fresh OSO4 diluted from the 4% water stock solution stored in the glass ampoules under argon in the dark at +4oC. At such conditions OSO4 is stable for at least a year even longer. I had some experience to store 2% OSO4 in 1x PBS at -20oC. I used NUNC 2 or 10 ml Cryo Tubes with (probably) silicone seal ring. At -20oC OSO4 vapors do not penetrate those tubes. To be double sure, I stored NUNC tubes in the 15 ml regular tissue culture polypropylene (?) tubes sealed with PARAFILM. At such conditions I stored OSO4 solutions in our Lab's refrigerator without any problems for a few months. As my knowledge, storing OSO4 in PBS is not a good idea. Frozen 2-4% water solutions (to be diluted 1:1 with 2x PBS for using) can be used instead with great success. Such stocks in full tubes (minimal amount of air) are stable for about half of year, even longer. Of coarse, all OSO4 solution must be store in the dark.
Good luck, Sergey
} Date: Mon, 25 Oct 1999 09:33:47 -0500 } From: "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com } Subject: Storage of Osmium tetroxide } To: Microscopy-at-sparc5.microscopy.com } X-MIMETrack: Serialize by Router on UNMCNOTES/Servers/UNEBR(Release } 5.0.1b|September 30, 1999) at 10/25/99 09:33:54 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
We have some users who are interested in using our new LEO 912 EFTEM for looking at meteorites. They are interested in purchasing an ion mill for thinning their samples. I would like to invite information and opinions from anyone regarding the current mills, and I would especially like to hear from vendors; I found many of their web sites had reply forms that didn't work!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
We have users of our Hitachi S-800 FESEM who are interested in a couple of accessories (and are willing to pay for them!). I would like to collect info and opinions about backscattered electron detectors and about cryostages for this instrument. Vendors are welcome to reply.
Please reply offline; I will relay messages to any other interested parties offline as well.
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Does anyone use this type of processor to embed tissue into Embed 812? We're having trouble with our stack of rings sticking in the transfer vial almost every time the machine moves the stack from one reagent vial to the next. We've tried new rings and vials and made sure the stack was straight. We're afraid to process overnight because of the hangups. There doesn't seem to be any pattern as to when the stack gets stuck. Any suggestions?
} Dear List, } Is there any problem with fixing and embedding macrophages using } standard protocols (GA,etoh,propylene oxide, P.O.parts epon etc), because } of enzymes etc on the surface. We seem to have fix/infiltration problems. } } Mike D. -
You'll have to be a bit more detailed if you want something more specific than a long lecture on basic procedure. Have you prepared macrophages successfully in the past with the same protocol? What kinds of problems? How old are your "epon" components? Do you use DMP-30 or BDMA as the accelerator?
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Can anyone suggest a source for low viscosity, high temperature epoxy adhesive, good to better than 600 F? I'm told that Epon828 will do the trick, but have no idea who makes it or where to get it.
Rick: Parafilm will not seal well enough and in any case, the oxygen within the vial will be trouble. As most of us have experienced, Os vapour penetrates plastic caps when stored in a freezer. I think that your options are another fridge or only purchase made-up osmium in small vials for use without further storage after opening. The commercial solutions are totally sealed in glass and stored under N2. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, October 26, 1999 12:34 AM, "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com [SMTP:"rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com] wrote: } } Hello All } This is probably one of those questions where everyone has there own } version but is 1% and 2% OsO4 in phosphate buffer solutions stable at cool } (20-22 C) room temperature? We want to store the double container- para } filmed solutions in the hood so we can do away with an old refrigerator but } the the only other refrigerator for the use of buffers and glutaraldehyde } chemicals is shared with an immuno-histotech that logically does not want } the osmium in there at all. } SPI alludes to keeping the diluted stock refrigerated, but the MSDS's we } have states room temp or cooler. Our safety people don't know except what } the MSDS says. } Thanks ahead for any help. } P.S. Does CAP regulations have any additional requirements? } } Rick Vaughn } rlvaughn-at-unmc.edu }
W. Erasmus wrote: ================================================= Can anyone recommend a diamond knife suitable for obtaining thin sections of Iron catalyst particles ? I currently have a Microstar SU knife with 2 mm length and 45 degree angle. After using it only once on the catalyst powder embedded in spurr's resin, I found that the edge was already scratched. Did I choose the wrong knife for this kind of work ? ================================================== We have been cutting "hard" materials, including catalysts of all types for nearly thirty years. Taking a new pristine diamond knife from any manufacturer, after the first "slice", there will be some striations, on these kinds of samples. And the more you section, the more will be the number of striations. Remember if just "touching" the edge with your finger can damage the edge, this should not be all that surprising, especially if what is being cut is iron powder.
Even before we entered the business of offering the knives ourselves, we had the experience of trying out knives from all of the major manufacturers and never saw any significant difference in the wear rate, on these kinds of samples, vendor to vendor, everything else (e.g. knife angle) being equal.
The real question however, has to do with just how deep are these striations and will they be sufficiently profound to ruin the chances of getting good sections. To be able to cut these kinds of samples with minimal creation of striations which are of minimum size is something that comes only from experience, technique, the samples themselves and how they are embedded, and perhaps even the ultramicrotome being used. You will get the best block for cutting if the samples are vacuum embedded. This won't reduce the wear on the knife edge, per se, however, it should enable you to get acceptable sections faster by using fewer knife "passes", and therefore resulting in less wear on your knife. Another hint would be to use a "harder" resin, such as SPI-Pon™, 812 or some of the other "Epon substitute" resin formulations.
Chuck
Disclaimer: SPI Supplies distributes the SPI Materials Science Diamond Knives and also sections these kinds of "hard" samples, for others, as a service for a fee.
============================================
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.spi.cc ######################## ============================================
After years of keeping osmium tetroxide solutions in a refrigerator I = finally tried another method. For over a year now, I have been keeping = aqueous solutions (up to 4%) at RT in a chemical hood, in a closed glass = bottle with no noticable problems. I also have a clean 'fridge.
If you are going to keep the aldehydes in a 'fridge, then make sure they = do not share with immunochemicals or tissue culture supplies. What = happens with the osmium will also happen with the aldehydes, they just do = not turn the sides of the "fridge black. Seriously, the aldehyde fumes = have the potential to fix protein solutions that are stored close by. = After all, that is what we want them to do.
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org
I am planning on studying microscopy as a hobby and for the education of my children. To that end, I just purchased a used American Optical microscope and I am looking for any and all information on this manufacturer and the history of their instruments. I haven't taken possession of the microscope yet and I understand that it does not have the original papers/instructions for care, etc. with it.
It is a binocular microscope which I believe is a Series 10. It has a mechanical stage, under-stage condenser, a reverse terret with 4 objective lenses (4x, 10x, 45x, and 100x), and 15x wide-field eyepieces. It also has a #1051 power supply for the illuminator.
If there are any knowledgeable members of the society out there who can give me some information on this piece of equipment and its history, I (and my daughters) will be very grateful.
Those interested in the history thread ought to note the book
"Picture Control" - The Electron Microscope and the Transformation of Biology in America, 1940 - 1960.
By Nicolas Rasmussen
Stanford University Press, Stanford, CA 1997
ISBN 0-8047-2837-2
Nicolas is now a lecturer at UNSW in Sydney, where I chanced on his excellent book.
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales Sydney, Australia. Phone +612 9385 6383 Fax +612 9385 6400
Those of you who are curious about the new toy microscope should read the review by the computing editor of the San Jose Mercury News (Silicon Valley's daily paper) at http://www.mercurycenter.com/svtech/computing/docs/ml102499.htm . He says that it has a CMOS chip, 288x352 pixels, & that the image is very high contrast. He also says that it's difficult to focus.
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I'm not sure about the usefulness of ion mills for meteorite work. We do quite a bit of this, and I find that FIB preparation is the best. Certainly with brittle samples you want to minimise polishing & mechanical thinning if possible, and the FIB allows us to cut a membrane wherever we want. However, it depends on the size of your sample and what information you require. We are usually concerned with micrometeorites collected from flight experiments or from Antarctica.
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Monday, October 25, 1999 11:44 PM To: Microscopy Listserver
Hello, all-
We have some users who are interested in using our new LEO 912 EFTEM for looking at meteorites. They are interested in purchasing an ion mill for thinning their samples. I would like to invite information and opinions from anyone regarding the current mills, and I would especially like to hear from vendors; I found many of their web sites had reply forms that didn't work!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} I'm looking for advice from anybody that has successfully fixed fungal
} yeast } cells in animal tissue. I am currently working with a thick-walled } yeast that } tends to crush in the tissue because of reasons unknown. I have } played with } the osmolarity of the fixative, as well as the dehydration time, both } to no } avail. The fixitives I have tried are trump's(4F:1G), and a 2.5% } glut. } fixitive. Some of the cells improved with the addition of sucrose to } the } fixitive, but there was still alot of crushed, hollow cells. If } anybody has } advice on the process or a good recipe for this kind of sample it } would be } appreciated. } } Eric Tarcha } Medical Technology } Michigan State University } tarchaer-at-msu.edu
I know nothing about fungi in animal tissue, but a lot more about fungi in plant tissue. Fungal cell walls seem to be less permeable to fixatives than plant cell walls and , of course, no comparison to animal
cells, also, young cells are easier to prepare than older stages. Are you sure that the yeast cells are alive and in good condition in the animal tissue?
Our prepartion protocoll for fungal infected plant tissue is: 2,5% buffered glutaraldehyde for 1 h, washing steps, then 0.5 -1% Osmiumteroxide, buffered, for 1 h at room temperature. Dehydration in ethanol or acetone, embedding in LR-White or Epon. We get better structural preservation if we dehydrate in the cold (PLT-Method): 1,5 h at zero (Grad Celcius) in 30% Ethanol, 1,5 at -20 in 50% Ethanol, overnight at -35 in 70% EtOH, 1,5 h at -35 in 100% EtOH. We cut very small samples if the tissue is difficult. Good Luck!
Anne Heller
---- Dr. Anne Heller, AG Elektronenmikroskopie, Institut f=FCr Botanik (210), Universit=E4t Hohenheim, Garbenstra=DFe 30 D-70593 Stuttgart
Attention List Members: RONTEC USA, Inc. is presently conducting a search for a Product Manager for its X-ray Microanalysis Group located in Acton, Massachusetts. If you have extensive SEM and EDX experience, are a US citizen and meet the job requirements of this position and wish to apply, please send a personal resume' and letter to: RONTEC USA, Inc.20 Main StreetActon, MA 01720Attn: Human Resources Written responses only. To view the job description, please go to: www.rontecusa.com “Employment Opportunities”.RONTEC USA is a subsidiary of the premier German EDX system manufacturer RONTEC AG. The growing RONTEC Group specializes in EDX detectors and analyzers of advanced technology, microanalysis and X-ray imaging software for SEM and TEM.
MAMAS will be meeting at NIST on Thursday, November 4, 1999 from 10:15am until 3:00pm. There will be coffee and doughnuts and two speakers. All are welcome. See below for details.
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
ANNOUNCEMENT ------------
Mid-Atlantic Microbeam Analysis Society (MAMAS) Meeting
at the National Institute of Standards and Technology (NIST) Gaithersburg, MD
on
Thursday, November 4, 1999 10:15am -- 3:00pm
Lecture Room E, Administration Building
10:15am: Coffee and Doughnuts
10:30am: Scott Walck, PPG Industries "The Small Angle Cleavage Technique"
Noon: Lunch
1:00pm: Joseph R. Swider, NIST Microanalysis Research Group "Micro X-ray Fluorescence of Particles Using a Laboratory X-ray Tube and Polycapillary Optics"
directions to the NIST campus can be obtained from the NIST website at: http://www.nist.gov/public_affairs/maps/nistmaps.html
for more information, contact John Henry Scott (NIST): (301) 975-4981 (301) 417-1321 FAX email: johnhenry.scott-at-nist.gov
Hi everyone. I am searching for an old part out of a Philips EM 400. The backing valve to the Diff pump and the buffer tank has failed. The bellows in it broke. It was originally made by Edwards. It is a Pipeline Valve Model PV 10. A P is also indicated behind the label PV 10 but stamped in not printed on. It is a small, simple Pneumatic straight valve with a bellows.
I have contacted Edwards and of course they don't have or make these anymore. The problem replacing this is the fittings or connections to it. They are of the large thread type(sorry don't know the proper name) not the KF type fittings that are commonly used now. I understand that the old old 400s used a different valve assembly. This is a "newer" 400 relatively speaking of course. I am hoping that someone has parts off an old 400 laying around that they would be willing to part with. My other course of action is to replace this valve with one readily obtainable which in turn will cause me to change other parts connected to this. Thanks everyone.
Joel McClintock U of Kentucky (606) 257-1242 jmcclin-at-pop.uky.edu
The rationale for storage of osmium tetroxide at 4 C has everything to do with oxidation and reduction. In fact, I never store osmium tetroxide in buffer. I keep a stock of 4% osmium tetroxide in distilled water at 4C, dilute to desired strength in appropriate buffer (only enough for the immediate processing) at time of use. Any buffered osmium tetroxide has a tendency to reduce to osmium dioxide and turn black, even at 4 C. Osmium tetroxide, including the stock will reduce fairly rapidly at RT (even 20-22 C). I know it is a bother, but we have managed to minimize the contamination of the fridge by double or triple containment (place the stock bottle inside one or two other larger containers). Hasn't completely stopped the problem, but it does minimize the general blackening of everything.
Roger Moretz Dept of Toxicology
} -----Original Message----- } From: "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com } [SMTP:"rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com] } Sent: Monday, October 25, 1999 10:34 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Storage of Osmium tetroxide } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello All } This is probably one of those questions where everyone has there own } version but is 1% and 2% OsO4 in phosphate buffer solutions stable at } cool } (20-22 C) room temperature? We want to store the double container- para } filmed solutions in the hood so we can do away with an old refrigerator } but } the the only other refrigerator for the use of buffers and glutaraldehyde } chemicals is shared with an immuno-histotech that logically does not want } the osmium in there at all. } SPI alludes to keeping the diluted stock refrigerated, but the MSDS's we } have states room temp or cooler. Our safety people don't know except what } the MSDS says. } Thanks ahead for any help. } P.S. Does CAP regulations have any additional requirements? } } Rick Vaughn } rlvaughn-at-unmc.edu }
Charles Brown wrote: =================================================== Can anyone suggest a source for low viscosity, high temperature epoxy adhesive, good to better than 600 F? I'm told that Epon828 will do the trick , but have no idea who makes it or where to get it. =================================================== M-Bond® 610 is a two component epoxy-phenolic resin adhesive system that, according to the manufacturer, when properly cured, has the following operating use ranges:
Short Term: -452°to +700°F (-269°to +370°C) Long Term: -452°to +500°F (-269°to +260°C)
More information can be found on the SPI Supplies website given below. Many persons asking for an epoxy seem to have been able to use the epoxy- phenolic system. SPI of course is a "source" for M-Bond 610 adhesive.
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.spi.cc ######################## ============================================
Dear all, About a week ago a posted a question regarding color CCD camera's for LM to this list. I promised than to send a summmary of the responses and here it is. I have also included a message on the same subject from the confocal e-mail list. To start with the original question is repeated here. Again many thanks to all who responded.
Original message:
We are in the stage of evaluating color CCD cameras for light microscopy use and would be very grateful for information and suggestions. I know this is a recurrent issue on this list and therefore I went trough the list archives. The most recent discussion I found was from a year ago and since there are quit some developments in this field, I guess there must be new information around.
The camera we are looking for will become part of our general light microscopy facility and will function in a multi-user / multi-application environment. It will be mainly used for imaging of histochemical and immunocytochemical stained tissues and cells -including imaging for quantitative image analysis-, but also for imaging of fluorescent signals in fixed or living cells and tissues grown on either glass or in plastic dishes. The camera should hence be suited for imaging in different modes: normal bright field, phase contrast, DIC and/or modulation contrast, dark field, epi-polarization and (to a lesser extend) also for fluorescence imaging but not for low signals.
The operation of the camera should be user friendly; most importantly it should offer a "fast focussing mode" (~ 10 frames a second) in either b/w or (preferably) color on the whole image or part thereof.
Resolution should at least be 1400x1000 real pixels (no color or pixel interpolation).
Whe now all this is very demanding and rules out already a lot of cameras. We concluded that what we need is a 36 bit RGB cooled camera with an interline transfer CCD and liquid cristal color filter. Are we correct in this? What are the advantages/disadvantages of this type of camera as compared to "normal three chip color CCD cameras"? Has anyone already used these types of cameras (three chip and interline single chip) in a setup as described above. More specific: any user comments on the CoolSNAP from Roper Scientific? Has anyone already seen the new SPOT RT from Diagnostic Instruments and can comment on it? Suggestions for other cameras?
Replies:
Is 1400 x 1000 pixels sufficient?
Check out these URLs for good info on different models and the DVC 1300 series camera.
http://www.epixinc.com/epix/
http://www.dvcco.com/dvc_1300.htm
gary g. ----------------------------------------------------------------------- Hi there,
I have evaluated the following:
1. use of digital input to computer requires frame grabber.
2. affordable frame grabber takes PAL or S-VIDEO input
3. such signals have maximum 700 x 500 pixels (or a couple of pixels more)
4. ccd camera does not have to produce too much more quality than that
5. single chip camera technically sufficient for still images
6. half-inch chip sufficient and technically matching image size projected from microsope through c-mount
7. microscope needs c-mount adapter to fit standard c-mount bearing of standard ccd camera
8. c-mount adapter with double oculars costs more than ccd camera
9. high resolution pictures are very nice, only the 1.6x enlargement has
problems because camera picks up slight irregularities in lighting not noticeable before
the first camera seller convinced me to spend much more money. anyway, we spent about 1800 sfr on the camera, and about the same amount for the leica c-mount adapter. i got a sony 1/2 inch single chip ccd-camera with pal output (color), and a ix-turbo-tv card (pci card for macintosh) for the video input along with a c-mount adapter for the microscope. the whole set works beautifully, except with things that this microscope can not do, such as producing high quality low magnification pictures.
the input i get is good enough for regular frame grabbing purposes, and perfect for the work that we do. we use differently stained light microscope with occasional polarization filters.
maybe this helps, good luck, wolf schweitzer md bern switzerland
Wolf Schweitzer wschweitzer-at-access.ch -------------------------------------------------------------------- Hello, My name is Robert Hocher and I work for a dealer sells Photometrics "coolSnap" and the SPOT-RT camera's. I also sell Hamamastu, Princeton, Optronics, and other HR digital camera's. What is your preference is operating system's Mc or PC Windows?? The CoolSnap is a good entry level
camera for Brightfield and very bright fluor..THe "RT" is just now being
released and I have no experience with it yet! I recommend staying with the Diagnostic Instruments SPOT-2e OR the Photometrics Sensys Color with
the enhanced Kodak KAF1401E chip. (Around $12500.00/each)Both camera's very good!! If you have alittle more money, than the Princeton Instruments MicroMax 5MHZ camera for $19500.00 is a very good choice with video output (25FPS). I can provide a quote to you, including image
analysis and digital camera solution if you want. I have prepared a list
of camera's I can forward as well! It shows QE% and specifications of alot of different cameras made today. What software are you using??MetaMorph/Optimas/Image Pro???...Phone 512-331-6685, Robert (rhocher-at-swbell.net) --------------------------------------------------------------------- Dear all, } We are in the stage of evaluating color CCD cameras for light microscopy
The LCD filter system you describe below uses a black and white ccd and the filter is used to achieve color by merging the 3 RGB images acquired separately.
} } use and would be very grateful for information and suggestions. I know this } is a recurrent issue on this list and therefore I went trough the list
} archives. The most recent discussion I found was from a year ago and since } there are quit some developments in this field, I guess there must be new } information around. } The camera we are looking for will become part of our general light } microscopy facility and will function in a multi-user / multi-application } environment. It will be mainly used for imaging of histochemical and } immunocytochemical stained tissues and cells -including imaging for } quantitative image analysis-, but also for imaging of fluorescent signals } in fixed or living cells and tissues grown on either glass or in plastic } dishes. The camera should hence be suited for imaging in different modes: } normal bright field, phase contrast, DIC and/or modulation contrast, dark } field, epi-polarization and (to a lesser extend) also for fluorescence
} imaging but not for low signals. } The operation of the camera should be user friendly; most importantly it } should offer a "fast focussing mode" (~ 10 frames a second) in either b/w } or (preferably) color on the whole image or part thereof.
The only way you can have a color preview at 10 frames per second with a digital camera is with a color ccd. All color ccds use interpolation to form the final image since individual pixels are masked with color filters. Color ccd cameras will never achieve the quality of image nor sensitivity you can get from a black and white ccd using LCD or glass filters to get color.
} } Resolution should at least be 1400x1000 real pixels (no color or pixel
} interpolation).
All the high resolution interline cameras except for the Spot RT use the Sony ICX-061 ccd. The ccd size of the ICX-061 is 1317X1035 in both black and white and color versions. One thing to remember here is that for a given magnification, a larger ccd does not give you any better image quality, all you get is a larger field of view.
The Spot RT uses a new ccd from Kodak and none of the cameras using that ccd have been released. Neuroscience USA in Miami will probably debut several cameras using that ccd.
} } Whe now all this is very demanding and rules out already a lot of cameras. } We concluded that what we need is a 36 bit RGB cooled camera with an } interline transfer CCD and liquid cristal color filter. Are we correct in } this? What are the advantages/disadvantages of this type of camera as } compared to "normal three chip color CCD cameras"?
The only cooled 3ccd cameras I am aware of use three 1/2 " ccd and are analog cameras with digital our. Analog cameras are limited to 8 bits per channel or 24 bit color. If your fluorescence requires more than one second exposure, you will need a cooled camera at least to 20 degrees C below ambient.
} Has anyone already used } these types of cameras (three chip and interline single chip) in a setup as } described above. More specific: any user comments on the CoolSNAP from
} Roper Scientific? Has anyone already seen the new SPOT RT from Diagnostic } Instruments and can comment on it? Suggestions for other cameras? } Please send me your comments and suggestions "off-line". I will be hapy to } send a summary to the list.
We have not yet seen the Spot RT. This may be a good camera. We have seen the Optronics Magnafire (www.optronics.com) and can highly recommend this camera. The two packages we strongly recommend are: 1)Cooke Sensicam (www.cookecorp.com) with Image-Pro Plus from Media Cybernetics and the CRI LCD filter system. 2)Hamamatsu Orca with Metamorph from Universal Imaging and the CRI LCD filter system.
But, if you want the best image quality for your money, check out www.apogee-ccd.com and remember that the faster your camera reads out, the more noise in your image.
Hope this helps. Shane Collins Imaging Specialist Scientific Instruments
This mail raised additional questions, which follow now together with Shane Collins' respons:
} The only way you can have a color preview at 10 frames per second with a digital } camera is with a color ccd. All color ccds use interpolation to form the final } image since individual pixels are masked with color filters. Color ccd cameras will } never achieve the quality of image nor sensitivity you can get from a black and } white ccd using LCD or glass filters to get color. The Magnafire form Optronics and the Spot RT from Diagnostic instruments do "promise" this in their brochures and without pixel interpolation. Or didn't I read between the lines? Do you have information that these camera's can not fullfill what is promised? } We have not yet seen the Spot RT. This may be a good camera. We have seen the } Optronics Magnafire (www.optronics.com) and can highly recommend this camera. Could you tell me a bit more why you highly recommend it? Any price indication? } The } two packages we strongly recommend are: 1)Cooke Sensicam (www.cookecorp.com) with } Image-Pro Plus from Media Cybernetics and the CRI LCD filter system. As I understand it, the SensiCam color does work with color interpolation (R, G and B pixels). Am I correct? Price indication? } 2)Hamamatsu } Orca with Metamorph from Universal Imaging and the CRI LCD filter system. I did not think about these camera's because to me they are the "top of the line" and consequently they are very expensive. Am I wrong on this? } But, if you want the best image quality for your money, check out www.apogee-ccd.com } and remember that the faster your camera reads out, the more noise in your image. Do you refer to the KX 85 camera (or any other camera of the KX series). Could you tell me a bit more why you think this camera gives the best image quality for our money?
Shane Collins answers to my additional questions: } The Magnafire form Optronics and the Spot RT from Diagnostic instruments do "promise" } this in their brochures and without pixel interpolation. Or didn't I read between the } lines? Do you have information that these camera's can not fullfill what is promised? The Magnafire uses a motorized filter wheel to change the glass filters. The speed of the filter wheel does not allow for the quick change necessary to change filter positions with enough speed to get multiple frames per second. Or at least that is what I experienced with the beta version camera. However, I may be incorrect in regards to the Spot RT. The LCD filters are capable of changing color fast enough to allow for 20 frames per second provided the computer can keep up. Cambridge Research (www.cri-inc.com) makes LCD filters that we use for variety of cameras. They have filters for black and white video cameras that give color preview in close to real time. Since we still need to take three pictures and merge them together to get a color picture from a black and white ccd, it makes sense that the frame rate for color would be 1/3 the frame rate of the black and white. That is unless Diagnostic Inst. have developed some camera electronics not previously available. Two comments on frame rate and LCD filters. A frame rate of 5, 10, or 20 frames per second is assuming that you have enough light to fill the electron wells to a level where signal to noise is high. In microscopy that is bright field mode most often. During fluorescence, the frame rate is exposure time plus read out time. With non intensified ccd you can expect the exposure time for fluorescence to range between 500 miliseconds to several seconds. The exposure time is clearly the limit to your frame rate. LCDs for RGB acquisition are only 40% efficient in light transmission and are not very useful for low light applications such as fluorescence.
Therefore, we usually take black and white images in fluorescence and colorize them with look up tables. The glass filters in the Magnafire transmit much better and can be used to acquire real color images in fluorescence. This feature could be quite useful in an auto fluorescence sample. } Could you tell me a bit more why you highly recommend it? Any price indication? The price should be around $12500 and uses a standard C mount so no expensive adapters are required. The Magnafire has a fast preview, is extremely easy to use in fluorescence, has built in coloroizing software, and is hot plugable so you can connect the camera to any computer that has a firewire port, turn it on and start taking pictures. No computer cards are required so you can connect to a laptop directly. } As I understand it, the SensiCam color does work with color interpolation (R, G and B pixels). Am I correct? Price indication? No, The SensiCam is a black and white ccd and you would merge three monochromatic images together to get color. There is no color interpolation with this method. Each pixel in each of the three images has its value express in the final image. The only adjustment is in color balance. } } 2)Hamamatsu } } Orca with Metamorph from Universal Imaging and the CRI LCD filter system. } I did not think about these camera's because to me they are the "top of the line" and } consequently they are very expensive. Am I wrong on this? No! you are quite right. The Sensicam with CRI and IPP will cost $26k. The Orca with CRI and Metamorph will cost $25k. But, with either of these systems you will have full image analysis capability and the Orca combination could be upgraded through software to do intracellular calcium measurements. } Do you refer to the KX 85 camera (or any other camera of the KX series). Could you tell } me a bit more why you think this camera gives the best image quality for our money? Yes, the KX 85 uses the same Sony CCD found in the Sensicam and Orca and therefore has the sensitivity and QE as those expensive cameras. And it is cooled to a lower temperature than the Orca. If you were only interested in fluorescence, the KX85 would work fine by itself for $6700. To add color you need the $3000 color filter option. But, your price is still under $10,000. The only negative to the KX85 is in the preview speed. You use the KX85 more like you use a film camera rather than a video camera. We sold KX 85s to UCLA, UCSF, Cal Tech, City of Hope, and many biotech companies, perhaps 14 so far this year in California. In summary, the best image quality for the least amount of money is probably the Cool Snap at $5000. The best image quality for the money is probably the KX85 at $10,000. The best all around camera for ease of use and multi user labs for fluorescence and brightfield is either the Magnifire or the Spot RT. Until I have the Magnifire in a demo next to the Spot RT, I won't know which is better. --------------------------------------------------------------- My colleague, Bill Delaney sent me your email message. My name is Harris Miller. I left Polaroid one year ago and now work with Bill in making microlenses for many interesting companies. Before leaving Polaroid, my
responsibility was color filter arrays for CCDs and CMOS image sensors. I was also part of a group that has built and is in the process of building several digital microscope cameras. Currently, I believe that Polaroid has the closest to what you want. I believe that Polaroid and Phillips are the only companies making "frame transfer" CCDs. Everyone else makes "interline transfer" CCDs. There is no difference in image quality. Everybody also uses color filter arrays (CFA) under the $120,000 range. The reason for this is otherwise, you have to optically cement three different CCDs to within 2.5 micron accuracy on a rather expensive piece of glass. However, Polarid uses linear filters. This means there is NO color interpolation in the vertical direction. Therefore, a true megapixel sensor (1,000,000 active pixels) produces file sizes in two different modes: 1.4 mg and 5.5 mg. We also completed a thermoelectrically cooled digital microscope camera which is capable of extremely long exposures. I don't know if this camera meets all of your needs, but I wish I had one here in our cleanroom.
Try www.polaroid.com website. Hope this helps.
Harris Miller 704 887 3155 ----------------------------------------------------------------- We demo'd the new Spot RT camera last week. It was really quite impressive. The specifications are on the web at: http://www.diaginc.com/spotrts.htm It is a supstantial improvement over the older Spot and Spot-2 cameras in that focusing is much closer to real-time and on the whole image...in B&W or color. Capture is also quite fast. There is substantial ability to control all camera features regarding exposure, gain, color balance, etc. It also is available for either MAC or PC. It is also a true 12-bit camera with resolution at 1520 x 1080 optical resolution. Actual capture speed depends on computer, video card, sample brightness, color or monochrome, and final resolution but can reach 12 frames/sec on color
and 19 frames/sec for monochrome. Price ranges from about $7500 to $12500 without adapter tubes for specific microscopes and a computer. Although
venders are not likely to have the camera at the moment (the company rep
did our demo), they should get them soon. It is certainly worth the wait to test it out before making a purchase of another camera. I have also demo'd the CoolSnap. It is a very nice camera .possibly the best at the lower end of the cost spectrum. (approx. $6000+ adapter tubes, etc.) Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------------------------------- Sounds like you may have already ruled out our choice, the Pixera. It will do up to 1280x1024, but is a little slower in that mode. I understand the technology is not interpolation, as such, but rather some shifting of the optics. Focusing is reasonably fast under the version 2.0 software.
We chose it since we are a general purpose materials lab and wanted a versatile camera. Yet we do not have so much demand for it to justify a large expenditure. If you are doing serious imaging, you probably need to more extensive cameras and you might as well get as much as you can now afford. ---------------------------------------------------------------- We have had excellent results using an Optronics DEI-750 CCD camera with a FlashPoint 128 video frame grabber for both fluorescence and bright field images. I understand the newer CCDs are even better and less expensive. The Optronics cameras are usually distributed via your Nikon and/or Olympus rep. Although I have not used an Optronics MagnaFire digital ccd camera, I have heard great comments about it and would definitely request a private demo. Specifications can be obtained at: www.optronics.com
Best regards, Robyn Robyn Rufner, Ph.D. Director, The Center for Microscopy and Image Analysis Ross Hall, Suite 406 Adjunct Associate Professor, Anatomy and Cell Biology THE GEORGE WASHINGTON UNIVERSITY 2300 Eye Street, N.W., 431 Ross Hall Washington, DC 20037-2337 Voice: (202) 994-2881 Fax: (202) 994-8885 Internet: anarrr-at-gwumc ----------------------------------------------------------------- Concerning using 6 bit cameras -- 6 bits are ok if you take full advantage of those 6 bits, but they leave you no room for error, and too little dynamic range.
Within this thread, someone asked about the DVC-1300 camera. I apologize
for not having the original post. In any case...
I have installed a DVC-1300C (the color version of the camera -- it also is available as a monochrome imager). The camera uses a Sony 1300x1000 pixel (give or take a few) CCD. It is a digital camera, produces a 12 bit/color output, and they are straight enough to admit that only 10 bits are meaningful. It is uncooled. You get up to 12 frames/sec.
Ours is coupled to an Epix digital frame grabber. The software is decent, and they have TWAIN and Image Pro drivers. The software control panel supports various integration modes and also fast shuttering (it is a line transfer CCD, so there is no need for a mechanical shutter). We have no problem seeing immunofluorescence and FISH. And this is with the color imager. The mono version will be much more sensitive. Integration times up to a few seconds can be done, without too much dark signal, even through it is uncooled. Moreover, you can raise the gain to focus and get a fast image, and then lower the gain and integrate when you like what you see. We like it, and bought a second one.
I believe that in most situations it will be more than adequate, although sometimes a cooled CCD will certainly do better.
However, if you are mainly doing fluorescence, then a mono imager is better than a color imager with color filters built into the CCD. You will find that in the price range of the DVC-1300 w/frame grabber (about $7000-$7500), you can find cooled monochrome imagers (check out Apogee systems cameras). If you want to add color capability later, many camera manufacturers let
you add on a CRI liquid crystal tunable filter (LCTF) to "convert" their
camera to acquire color images. If you want something inexpensive that can make excellent images, check out Electrim cameras (they are only digital). But then be prepared to write some software to take full advantage of those cameras. There is a paper in Optical Engineering from at least 5 years ago (sorry, I dont have the ref handy) which shows that in many cases successive digital integration of images from a conventional video rate camera can work as well as on-chip integration. It depends on the readout noise of the camera system. But, if you dont sync on the pixel clock (and cheap cameras often dont have that available), then you get pixel jitter. I still think you are better off with a digital camera, if you have the budget.
--aryeh Aryeh Weiss | email: aryeh-at-optics.jct.ac.il Department of Electronics | URL: http://optics.jct.ac.il/~aryeh Jerusalem College of Technology | phone: 972-2-6751146 POB 16031 | FAX: 972-2-6751275 Jerusalem, Israel | ham radio: 4X1PB/KA1PB ---------------------------------------------------------------------------------
END OF REPLIES --
Lauran C.J.M. Oomen Phd - Manager Digital Microscopy Facility Department of Biophysics The Netherlands Cancer Institute voice: +31 20 512 1887/1889 Plesmanlaan 121 fax: +31 20 512 1893 1066 CX Amsterdam email: oomen-at-nki.nl http://www.nki.nl/nkidep/biofys/microscopy/digmiclo.htm
We have a BalTec RES 100 ion mill. I really like my machine after some of the initial bugs were worked out of it. It is very flexible to use and can be applied to work with SEM samples for both polishing and ion etching. There is a technique called slope cutting that allows layers to be exposed and beveled. I highly recommend it. Leica is the representative in the US. -Scott
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Monday, October 25, 1999 5:44 PM To: Microscopy Listserver
Hello, all-
We have some users who are interested in using our new LEO 912 EFTEM for looking at meteorites. They are interested in purchasing an ion mill for thinning their samples. I would like to invite information and opinions from anyone regarding the current mills, and I would especially like to hear from vendors; I found many of their web sites had reply forms that didn't work!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
Willem, We use only 55 degree angle knives for hard materials. I must say however that we take great care in minimizing the specimen particle size prior to embedding. We usually do this by extensive hand grinding with a small, hard, mortar and pestle. For me, I find that very hard materials that are likely to damage the diamond knife, will not grind smoothly or uniformly into a nice powder. This step is usually the point at which we make a decision to continue with the ultramicrotomy prep, or try something else. Another aid to microtomy of hard materials is the infiltration of the embedding resin into the structure of your material. If possible, we use LR White (hard) or similar acrylic resins because of the wetability that many materials have with these resins. LR White often gives us great infiltration without the aid of vacuum. For materials that require epoxy resins, I also use Spurr's resin first but always with vacuum infiltration; again try to match specimen hardness as appropriate. Another point, don't use your favorite knife for microtomy of hard materials. At least test on an old knife that is ready to go back for resharpening. I've had a few surprises over the years when I thought all was well, only to have a chunk of the knife to disappear! Good Luck, Brad BPAmoco
} ---------- } From: Erasmus, Willem (WJ)[SMTP:willem.erasmus-at-sasol.com] } Sent: Monday, October 25, 1999 9:10 AM } To: 'microscopy-at-msa.microscopy.com' } Subject: TEM Ultramicrotomy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopists } } Can anyone recommend a diamond knife suitable for obtaining thin sections } of } Iron catalyst particles ? I currently have a Microstar SU knife with 2 mm } length and 45 degree angle. After using it only once on the catalyst } powder } embedded in spurr's resin, I found that the edge was already scratched. } Did } I choose the wrong knife for this kind of work ? } } Sincerely } W. Erasmus } } Willem Erasmus } Snr. Scientist, Basic Catalysis Research } Sasol Technology } Tel : +27 +16 9604211 } Fax : +27 +16 9602826 } E-mail : willem.erasmus-at-sasol.com } }
} Hi everyone. I am searching for an old part out of a Philips EM 400. The } backing valve to the Diff pump and the buffer tank has failed. The bellows } in it broke. It was originally made by Edwards. It is a Pipeline Valve Model } PV 10. A P is also indicated behind the label PV 10 but stamped in not } printed on. It is a small, simple Pneumatic straight valve with a bellows. } } I have contacted Edwards and of course they don't have or make these } anymore. The problem replacing this is the fittings or connections to it. } They are of the large thread type(sorry don't know the proper name) not the } KF type fittings that are commonly used now. I understand that the old old } 400s used a different valve assembly. This is a "newer" 400 relatively } speaking of course. I am hoping that someone has parts off an old 400 laying } around that they would be willing to part with. My other course of action is } to replace this valve with one readily obtainable which in turn will cause } me to change other parts connected to this. Thanks everyone.
Contact Ron Veil at Veil Electron Instrument Lab. He has parts for the 400. He is at 650 952 3099, 173 Santa Inez Ave., San Bruno, CA 94066.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
} } I am planning on studying microscopy as a hobby and for the education of my } children. To that end, I just purchased a used American Optical microscope } and I am looking for any and all information on this manufacturer and the } history of their instruments. I haven't taken possession of the microscope } yet and I understand that it does not have the original papers/instructions } for care, etc. with it. } } It is a binocular microscope which I believe is a Series 10. It has a } mechanical stage, under-stage condenser, a reverse terret with 4 objective } lenses (4x, 10x, 45x, and 100x), and 15x wide-field eyepieces. It also has a } #1051 power supply for the illuminator. } } If there are any knowledgeable members of the society out there who can give } me some information on this piece of equipment and its history, I (and my } daughters) will be very grateful. } } Mike T. } maturco-at-aol.com
Mike -
Congratulations on your purchase and plans for the future! Your AO is a good, solid basic oldie. Fortunately for you, such scopes don't need model-specific instructions. The first book for you to purchase is Nachtigall, "Exploring with the Microscope"; see the MICRO bibliography (URL below) for ordering information. If you have further questions, please contact me.
Caroline Schooley Project MICRO 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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I have an old EDAX 9900 here at ANL, whose Display Monitor has bitten the dust and I'm looking for a replacement.
If anyone has one they would like to part with please contact me off-line ASAP. I have a visiting student here who needs to get some work done and is only here for 2 months.
This is a special RGB Monitor with BNC connectors that is driven off custom display boards in the 9900. The monitor Model # is 38-DO5IMA-XI, made by ElectroHome (who has since been bought out by Conrac). This model is nolonger manufactured and the suggested replacement from Conrac (with no guarentee that it will actually work) is $1300 with 6 week delivery.
I've been trying various standard RGB monitors without success because of the sync "frequency" used the graphics display board.
Any help or suggestions of alternate monitors that people have found will be appreciated.....
BTW, I've also contacted EDAX and they are also looking for sources of replacements. The 9900 system is ~ 10 years old so spares are not necessairly off the shelf.
Thanks in advance...
Nestor
================================================================== Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov ================================================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ==================================================================
The box said "This program requires Win 95/98/NT or better..." so I bought a G3 Mac
Dear Tina, I am fortunate to have both a GW backscattered detector, which is the solid-state type, and a Robinson, which uses a fluorescent detector. Both are on Hitachi W filament SEMs. They have different strengths and weaknesses. The Robinson is brighter and will give you a picture with more contrast at lower beam currents. The GW is more sensitive to atomic number contrast, and with careful tuning will show you things the Robinson cannot, although at a higher beam current. Since your S-800 is a field-emmission, it might be somewhat restricted in total beam current output, so that might influence your choice. I have no experience with cryostages. At 11:48 AM 10/25/99 -1000, you wrote:
} Hello, microscopists- } } We have users of our Hitachi S-800 FESEM who are interested in a couple of } accessories (and are willing to pay for them!). I would like to collect } info and opinions about backscattered electron detectors and about } cryostages for this instrument. Vendors are welcome to reply. } } Please reply offline; I will relay messages to any other interested } parties offline as well. } } Mahalo, } Tina } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
At 09:50 AM 10/26/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
[big snip]
As is the case in most areas, there are tradeoffs here. I need high quality images that I can directly capture to computer and will provide real time video imaging. I'd like to get them for $5. Can't be done.
I settled on two options. Sony DKC-5000 "Cat's Eye" and the DKC-ST5. These are SCSI interface cameras with RGB+sync outputs to a color video monitor for real time preview and focusing, composition, etc.
The 5000 is a 1/2" CCD with 440,000 pixels with an effective image area of 795 x 598 pixels. In memory mode, the image is reduced to 738 x 576 pixels. These are 8-bit pixels per RGB color plane.
The ST5 effectively doubles the pixel dimensions of the 5000. The cost of the older 5000 is about $8K while the ST5 is about $15K. Using stitching software, either camera will produce the same final image. It just takes more shots with the 5000.
The main issue is how detailed and large of a final image do you want?
Hi, I'm trying stain stomata without success. My samples are claryfied at hypoclorite. Someone could help me? Thanks, Rejane Pimentel Universidade Federal Rural de Pernambuco Brazil Rejane Magalh=E3es Pimentel Galindo Functional Plant Morphology Universidade Federal Rural de Pernambuco ggalindo-at-elogica.com.br Av. Boa Viagem, 6592/602 51130-000 Boa Viagem
Dear Richard, I have two references by John Kuo on removing stain precipitates:
"A simple method for removing stain precipitates from biological sections for transmission electron microscopy" by John Kuo, Journal of Microscopy, Vol.120,Pt 2,November 1980, pp.221-224. and "Forming and Removing Stain precipitates on ultrathin sections", J.Kuo et al., Stain Technology, Vol.56, No.3, 1981, pp.199-204.
Also a brief note by H.Mollenhauer and D.Morre, 1978 "Contamination of thin sections, cause and elimination", in Electron Microscopy 1978, Vol.II, pp.78-79. (edited by J.Sturgess et al, Imperial Press, Ontario).
There is more than one method in these articles depending on the type of contamination, so I won't try to make a suggestion. Susan
================================================================= Susan Belfry Electron Microscopy Unit email: belfry-at-unb.ca University of New Brunswick phone: 506-453-4887 Bag Service No.45111 fax: 506-453-3583 Fredericton, New Brunswick, http://www.unb.ca/emunit E3B 6E1 Canada =================================================================
Of course this begs the question of why you are looking to get rid of a year old FEG SEM?
} The University of Michigan Materials Science and Engineering } Department has the following machine available. } } } Hitachi S-800 FEG SEM with GW Electronics Microchannel Plate BSE detector } and Macintosh-based 4pi Analysis digital image acquisition system. } Instrument purchased new in 1988 and maintained under service contract } until Sept., 1999: $70,000. For further information call (734) 764-3357. } _______________________________________ } Anne E. Huber Ph.D., Instrument Analyst } Materials Science and Engineering Dept. } The University of Michigan } 2300 Hayward St. } Ann Arbor, MI 48109-2136 } ahuber-at-umich.edu } (734)764-3357 } _______________________________________ }
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
} Does anyone know of the method of removing lead citrate post stain } artifacts. I remember there is a protocol but can't seem to find it. } Richard Gardiner } Agriculture Canada
Richard, There are 3 references in the EMS catalog (pg 30). One listed is: Kuo, J., et al(1981). Forming and removing stain precipitates on ultrathin sections. Stain Technol., 56:199
hope this helps, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
We have two instruments now in excess and available (best offer): a scanning electron microscope ISI40, and a transmission electron microscope Philips400, PW6585. If you are interested please contact: Pat Ashbaugh (ashbaugh.11-at-osu.edu) or Tea Meulia (meulia.1-at-osu.edu).
Tea Meulia Research Scientist Head Molecular and Cellular Imaging Center OSU/OARDC 1680 Madison Ave. Wooster OH 44691
We have a male post doc from Germany who would like to share a room with someone during the MRS meeting in Boston--November 30 - Dec 3. Please respond directly to my address and I will give him the replies as they come in. Thank you.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
We too use P2O5, but at a rate of about twenty 500g bottles a year (We go through around 90 - 100 boxes of Kodak SO-163 in that time). Each time the desiccator is opened, we use just enough powder to cover the surface of a six inch petri dish with a thin layer. When I saw Rick used only two bottles in ten years made me think that we are using too much.
What is the usage like in other labs?
Joan MicroSci-at-AOL.com
In a message dated 10/27/1999 11:19:31 AM, rlvaughn-at-unmc.edu-at-Sparc5.Microscopy.Com writes:
} We have been using P2O5 for over ten years and I think we're on our second } bottle so the amount going into the environment is negligible (on our } part). We have used silica and calcium dehydrants and they did not work } as } well or last as long. } To increase the longevity of the dehydrant and aid in keeping the film } and } container dry we also used dry nitrogen gas to evacuate the desiccator, } which is also connected to the scope for camera chamber evacuation. On } a } identical scope and desiccator with out the nitrogen we have much slower } cycle times, and the desiccant is exhausted more quickly. } The P2O5 is nasty to work with though so I will be interested in seeing } what other chemicals people come up with. } } Rick Vaughn
Perchance does anyone have a good wet etch to remove oxide (SiO2) and leave an underlying titanium silicide film intact? We have tried BOE 7:1 HF (buffered with ammonium hydroxide) and the results were crappy. We have also tried [48%] HF and had reasonable success but timing is very, very critical here. Anybody have any other suggestions???
Thanks!!! John W. Staman LSI Logic, LLCO Analytical Services 1-719-573-3282 (VM) 1-719-577-3127 (Pg.)
We have a user who wants his own double-tilt specimen holder, preferably with a Be cup for EDS work, for a Philips CM30T TEM. A holder from the 400 series TEMs should work fine. Does anyone have a used holder they'd like to sell? Please contact me directly. Thanks.
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798 cook-at-aaem.amc.anl.gov
I have a CoFe2O4 film on an MgO substrate. I am looking for a way to remove the MgO (chemically, mechanically, etc.)and leave behind just the CoFe2O4 film intact. To date I have been able to mechanically polish away all but about 15 microns of the MgO. When the MgO gets thinner than this it tends to just cleave and break up, and then break the CoFe2O4 film as well. I would be very grateful for any suggestions.
Thanks in advance,
Mick Thomas
---------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
We have a problem with our grids often being hydrophobic, even when used less than an hour after glow discharging.
The carbon film is deposited on nitrocellulose coated grids inside a relatively new turbopumped evaporator, so we hope there is no hydocarbon contamination there. Our glow discharge unit is kept clean and has a foreline trap - again we hope no contamination. When we glow discharge we typically pump for about twenty seconds and glow discharge for ten to twenty seconds.
Anybody have a similar experience or any ideas?
Thanks, Joan MicroSci-at-AOL.com
In a message dated 10/27/1999 11:22:21 AM, jubu-at-uclink4.berkeley.edu writes:
} Yuhui Xu asked: } ================================================ } Two questions: } 1. Which vendor sells formvar + carbon coated grids which are also } glow-discharged } ========================================================== } Hi Yuhui, } We make our own grids, means carbon coat them & also glow them before } use.We had the same problem of grids getting hydrophobic after 2 weeks. Try } this it works,after you glow them store grids in the refrigerator till you } need it .If you do this the grids remain hydrophilic even after a month.Get } your grids out before you need them & when your are done stick them back in } the refrigerator. } } Reena Zalpuri } EM Lab } UC Berkeley } E-Mail jubu-at-uclink4.berkeley.edu
The P2O5 goes further and lasts longer if you mix it with vermiculite.
The vermiculite gives the P2O5 a greater surface area and avoids the problem of useful oxide getting trapped under the phosphoric acid layer.....
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales Sydney, Australia. Phone +612 9385 6383 Fax +612 9385 6400
We have two instruments now in excess and available (best offer): a scanning electron microscope ISI40, and a transmission electron microscope Philips400, PW6585. If you are interested please contact: Pat Ashbaugh (ashbaugh.11-at-osu.edu) or Tea Meulia (meulia.1-at-osu.edu).
Tea Meulia Research Scientist Head Molecular and Cellular Imaging Center OSU/OARDC 1680 Madison Ave. Wooster OH 44691
Subject: LWD objective lenses Hello all, I'm looking for LWD objective lenses with a WD} 9mm. The magnification should be {20 and the nA} 0.13. Can you please give me some more data of any microscope objective lenses? Thanks a lot, Kristiane ----------------------------------------------------------------------------= ---- ----- Kristiane Gans Leipziger Stra=DFe 44 ZENIT (Haus 65) Institut f=FCr medizinische Psychologie 39120 Magdeburg Germany Tel.: 0391 - 6117 - 106 =46ax: 0391 - 6117 - 103 Mail: Kristiane.Gans-at-medizin.uni-magdeburg.de Attachment converted: Zaluzec-0:Kristiane Gans.vcf (TEXT/R*ch) (00022DE0)
} We too use P2O5, but at a rate of about twenty 500g bottles a year (We go } through around 90 - 100 boxes of Kodak SO-163 in that time). Each time the } desiccator is opened, we use just enough powder to cover the surface of a six } inch petri dish with a thin layer. When I saw Rick used only two bottles in } ten years made me think that we are using too much. } } What is the usage like in other labs? }
Dear Joan, In the desicators where we use P2O5 we add ~ 1 Tsp/change (~10 g), and we change it once/day each day the film is changed. This works out to ~4 500 g bottles/year. Yours, Bill Tivol
I treat my grids for 10 minutes at 1.25 Amp with air leaking into a dp = system.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } {"MicroSci-at-aol.com"-at-sparc5.microscopy.com} 10/27 5:27 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html=20=
We have a problem with our grids often being hydrophobic, even when used = less=20 than an hour after glow discharging.
The carbon film is deposited on nitrocellulose coated grids inside a=20 relatively new turbopumped evaporator, so we hope there is no hydocarbon=20=
contamination there. Our glow discharge unit is kept clean and has a=20 foreline trap - again we hope no contamination. When we glow discharge = we=20 typically pump for about twenty seconds and glow discharge for ten to = twenty=20 seconds.
Anybody have a similar experience or any ideas?
Thanks, Joan MicroSci-at-AOL.com=20
In a message dated 10/27/1999 11:22:21 AM, jubu-at-uclink4.berkeley.edu = writes:
} Yuhui Xu asked: } =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D } Two questions: } 1. Which vendor sells formvar + carbon coated grids which are also } glow-discharged } =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D } Hi Yuhui, } We make our own grids, means carbon coat them & also glow them before } use.We had the same problem of grids getting hydrophobic after 2 weeks. = Try } this it works,after you glow them store grids in the refrigerator till = you } need it .If you do this the grids remain hydrophilic even after a = month.Get } your grids out before you need them & when your are done stick them back = in } the refrigerator. } } Reena Zalpuri } EM Lab } UC Berkeley } E-Mail jubu-at-uclink4.berkeley.edu=20
Double distilled water comes up with alarming frequency in the electron microscopy literature. It is commonly mentioned in immunocytochemistry techniques but also seems to be popular with some workers for all techniques, especially staining
My suspicion is that purity is important but consistency is even more critical, especially for more demanding work such as e.m. cytochemistry, immunocytochemistry, DNA spreading etc.
Does anyone know of any literature or personal evidence about the superiority of double distilled water (pyrogen-free) and high quality de-ionised (low pyrogen but high resistance/low conductivity) water or is this more dogma? Has there been a FAQ on this subject that I have missed?
thanks Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
We find that after two or three 30sec cycles of glow discharge in our Balzers Union CTA 010 our carbon coated grids are reasonably hydrophilic for about an hour. Even if sample droplets appear to spread well macroscopically there often small microscopic patches of hydro- phobicity. As a matter of routine we discharge again after an hour or so to promote uniform adherence. Don Gantz Biophysics Dept Boston Univ School of Medicine
A new and (I hope) simple question. We were lucky to borrow a CO2 critical point drying device, an older Balzer's CPD 020 for teaching purposes. It will stand in for an even older Bowmar 900 unit that used Freon which is no longer available. Part of the apparatus with the loaner is an in-line filter for CO2 that actually goes into the bomb for exchange. The filter is ostensibly to remove oil, water, and particulates from the CO2 which might compromise the CPD conditions or contaminate the specimen. The unit included is marked: Union Carbide high pressure filter, model SG 6140 which uses a removable cartridge which comes sealed in a ring pull can marked Union Carbide purifier cartridge part no. SG 6142. The holder is a well turned heavy duty brass cylinder which unscrews to allow the cartridge to be changed. The cartridge screws into the top of the holder and appears to be full of a desiccant resin bead of some sort.
Does anyone know of a source for the replaceable cartridges? Union Carbide's web page was no help at all and our purchasing specialists at the University haven't had any luck yet, either.
Thanks in advance for your help-
Regards, Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
One of our facilities has an ISI SX-40A SEM which is no longer needed. This microscope is operational, but has been out of service. It needs to be moved to a good home. For additional information, please contact: Gary Troyer Ferro Corporation Diamonite Division 453 W. McConkey Street Shreve, OH 44676 330-567-2145 troyerg-at-ferro.com
David Gnizak Ferro Corporation Technical Center 7500 E. Pleasant Valley Rd. Independence, OH 44131 gnizak-at-ferro.com
I may be looking for a used, but recent model, FESEM at a good price. A high vacuum model is fine. I need a motorized 5 axis stage and full computer control (Win95/98). Computer image capture is not a necessity but the scope must be able to accept the Soft-Imaging ADDA active/passive interface.
Thermally assisted FESEM is OK. A 4" chamber is highly desirable. Turbo pump is necessary. Reliable mechanical pump is required.
I would like to be able to use my Amray specimen stubs (3.1mm dia stubs, 12mm diameter).
The scope will also need to qualify for factory service/maintenance contract. I would plan on factory take down, crating and separate moving but factory re-install at my site (Sacramento, CA).
Dear Malcolm, As my knowledge, there are two types of water contamination, which important in EM and biochemical techniques: "organic" contamination (including the products of microbes degradation etc.) and CO2. Double distillation (preferably with quartz-distillator at the second stage) removes organic compounds from the water (especially if you add some potassium permanganate in the water). Fresh prepared distilled water (not necessarily "double"-distilled) does not contain CO2. Organic components are difficult to detect and they do not interfere with conductivity. Sometimes you may have very low conductive water still contaminated by phenols for instance. You may detect such contamination by checking UV spectra around 220-240 nm. From another hand, pure water exposed to the air for long period of time (couple of hours) will accumulate CO2 and becomes high conductive. Modern ion-exchange column water purification systems include the ion-exchange resins to remove inorganic components. To remove "organic" contamination activated charcoal were used. The resin itself may be a source of the "organic" contamination. Microbes or products of its degradation, also, may contaminate the tubing of the purification system down stream of the purification columns and sensors (therefore, this contamination does not detect by sensors). As for "Pyrogen/pyrogenity": people injected a couple milliliters of water into rabbit's blood stream and check the body's temperature. If the temperature is higher than normal - it mean that water is "pyrogenic".
The conclusion is: "cell culture grade" 18-20 MOhm/cm water from purification system (not bottled water) not necessarily "pure" enough for your particular application. It may contain some organic compounds including the product of microbes degradation, which may interfere with your protocol. Fresh double-distilled water in most cases is pure enough for most EM and biochemical applications. It, also, comes "sterile" because of boiling and "CO2 free". From another hand, I find that modern water purification systems provide water suitable for most EM and biochemical applications. If it is necessary, I remove CO2 by 20 min boiling or by bubbling of the He. Bottled "HPLC" or "Cell Culture" grade water from FISHER or any other biotech. Companies may be the solution, too.
Sergey
} Date: Thu, 28 Oct 1999 16:00:33 +0000 (GMT) } From: malcolm.haswell-at-sunderland.ac.uk (HASWELL Malcolm) } Subject: Water - to double distill or not? } To: microscopy-at-sparc5.microscopy.com (Microscopy) } Organization: University of Sunderland } X-Mailer: Microsoft Mail via PostalUnion/SMTP for Windows NT } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I recall in a previous discussion someone said that they avoided de-ionised water as a failing system added unwanted material to the water.
Dave
On Thu, 28 Oct 1999 16:00:33 GMT HASWELL Malcolm {malcolm.haswell-at-sunderland.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Double distilled water comes up with alarming frequency in the electron } microscopy literature. It is commonly mentioned in immunocytochemistry } techniques but also seems to be popular with some workers for all } techniques, especially staining } } My suspicion is that purity is important but consistency is even more } critical, especially for more demanding work such as e.m. cytochemistry, } immunocytochemistry, DNA spreading etc. } } Does anyone know of any literature or personal evidence about the } superiority of double distilled water (pyrogen-free) and high quality } de-ionised (low pyrogen but high resistance/low conductivity) water or is } this more dogma? Has there been a FAQ on this subject that I have missed? } } thanks } Malcolm } } Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Please advice me if there is any image processing and analysis system for SEM, Philips 500 series. Features needed: able to count numbers of predetermined objects / particles in a certain area and produce hard copy images.
Please advice me if there is any image processing and analysis system for SEM, Philips 500 series. Features needed: able to count numbers of predetermined objects / particles in a certain area and produce hard copy images.
Bill, why bother with that filter? If you get food-grade C02 (as is used to gas beer in hotels) no filter is required. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, October 29, 1999 8:25 AM, William P. Sharp [SMTP:wsharp-at-asu.edu] wrote: } } } Hello, Listers - } } A new and (I hope) simple question. We were lucky to borrow a CO2 critical } point drying device, an older Balzer's CPD 020 for teaching purposes. It } will stand in for an even older Bowmar 900 unit that used Freon which is no } longer available. Part of the apparatus with the loaner is an in-line } filter for CO2 that actually goes into the bomb for exchange. The filter is } ostensibly to remove oil, water, and particulates from the CO2 which might } compromise the CPD conditions or contaminate the specimen. The unit } included is marked: Union Carbide high pressure filter, model SG 6140 which } uses a removable cartridge which comes sealed in a ring pull can marked } Union Carbide purifier cartridge part no. SG 6142. The holder is a well } turned heavy duty brass cylinder which unscrews to allow the cartridge to } be changed. The cartridge screws into the top of the holder and appears to } be full of a desiccant resin bead of some sort. } } Does anyone know of a source for the replaceable cartridges? Union } Carbide's web page was no help at all and our purchasing specialists at the } University haven't had any luck yet, either. } } Thanks in advance for your help- } } Regards, Bill Sharp } William P. Sharp } Arizona State University } Dept. Plant Biology, box 871601 } Tempe, AZ 85287-1601 } Phone - (480)-965-3210 } Fax - (480)-965-6899
Fisrtly, many thanks to all those who replied to my query about negative scanners. We still haven't got around to deciding, but at least I have the information.
I would like to recommend a scanner book which I recently acquired for our Polymer Group here in Reading. I'm not suggesting you all rush out and buy it immmediately, but if your department at work is considering buying a scanner or needs training in using one to best advantage, then I would recommend this as a departmental purchase.
You can find out about it on:
http://www.scantips.com/
and the book version in particular on:
http://www.scantips.com/book.html
It is called "A few scanning tips" by Wayne Fulton (from Texas) and costs about $22 (US) + shipping. It tells you all sorts of things you might want to know about scanning, from the simple basics to all sorts of things about PC settings, etc. Although it is based mainly on examples from Microtek scanners, the principles are applicable to all scanners, both flat bed and film.
+------------------------------------------------------------------------+ | 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 have a two dessicator system for drying our films. One dessicator contains the driest plates to be used first and the other dessicator contains a second camera box of unused plates and previously opened packets of film. Each dessicator contains a thin layer of P2O5 powder in a standard sized petri dish. We check twice a week for degree of hydration and change when appropriate. Our dessicant consumption is four or five 500g bottles per year for 7,000 - 9,000 exposed films.
Don Gantz Biophysics Department Boston Univ School of Medicine
Hi, I have to sign service contract soon, and I have a question: Is it a usual practice (by manufacturer) to exclude EDS detector's windows from coverage, even as an option?
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Does anyone know where to purchase Kainic acid, formerly Sigma K-0250? I understand that it it off the market and one of my friends is very much in need for their study. Thanks, Linda Fox lfox1-at-wpo.it.luc.edu
As I understand it, Philips had used Media Cy's Image-Pro Plus for a short period of time. Our research (M&M '99) shows that it is still the Number One off-the-shelf software on the market. It will easily do what you ask.
Caveat: MME has no financial interest in this product.
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 07:00 AM 10/29/99 -0500, Budi Widagdo wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Many thanks to all for numerous replays to my question! Unfortunately there is no simple way to print... So, for now I will just use screen capture, and it is not a good way to use our Fujix Pictrography 3000 printer with 3800*2759 print pixels. I just hope that PGT will be more user friendly in future and release file conversion utility (to do it by myself will be too time consuming).
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] } Sent: Monday, October 18, 1999 10:34 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Imix PC pictures } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } ---------. } } } After installation and working a lot with my new EDS systems (Imix PC) } from PGT, I am now preparing publications. And - surprise, surprise, - } I have found out that I cannot print good quality pictures (on } high quality printer which is not connected to our network)! } } The file format of stored images, RAS with overlays, cannot be read by } other programs. Of course, some of them can read RAS, but overlays } (with the most important information) are lost. } } The only advice I've got from PGT was to capture images from } the monitor. } But then I will have files with much lower resolution than } initial ones, } and this is certainly bad for high quality printing. } } All (if any) advises will be highly appreciated!!! } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } }
do you need only processing or also acquisition form the microscope? Is passive acquisition sufficient, or do you need active acquisition? In any case, you may want to check out our web site for our ADDA II product and the analySIS software. We actually have a Philips 500 Series Microscope in our development facility, so I can gurantee that it works ;-).
If you tell me, where in the world you are located, I can also point you to the nearest contact.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Budi Widagdo[SMTP:BDWDGD-at-CENTRIN.NET.ID] } Sent: Friday, October 29, 1999 3:14:17 AM } To: Microscopy Society of America } Subject: Images processing and analysis for SEM } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear everybody,
Please advice me if there is any image processing and analysis system for SEM, Philips 500 series. Features needed: able to count numbers of predetermined objects / particles in a certain area and produce hard copy images.
I have a problem with my EDS detector installed with ESEM. I often use "wet" mode with water pressure up to 10 Torr. Detector is pretty cold (I think its temperature is about 12-15C) and it can lead to water condensation on its window. With time detector become colder and colder, and than - frosted. As a result I have had two window replacements in less than one year. Is this problem the usual thing for "wet" ESEMs? Are all new detectors colder than older ones (which are much thicker)?
Thanks,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
{smaller} A postdoctoral fellow position is available at the University of Illinois at Chicago for the development and application of novel STEM/TEM techniques to the analysis of metal-oxide interfaces and grain boundaries in metals. Research in the Interface Physics Group focuses on the use of atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Other related research programs within the group include ceramics, ionic conductors, high-Tc superconductors and semiconductor materials/devices, and it is expected that the successful applicant will work closely with the existing group members in these areas. =20
The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, Noran EDS, Gatan liquid nitrogen cooling stage, and Gatan heating stage; a VG HB601 Field-Emission dedicated STEM featuring a 2.2=C5 probe size and EDS and EELS capabilities; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 =46ield-Emission SEM with EDS; and a JEOL JXA733 microprobe. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, Fischione precision ion-mill, SouthBay plasma cleaner and Leica Ultramicrotome. The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with the Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. =20
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevant materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. =20 However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience.=20 UIC is an equal opportunity employer.
We have a very limited amount of protozoan sample which appear to have virus-like particles within the cells, but we need to know more conclusively if the particles are virus before we continue. Does anyone know of a method using EM to identify virus in tissue? We presently have material available and could run either a pre or post embedding technique.
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
At 08:00 AM 10/29/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From what I have researched, check out Soft-Imaging and their analySIS and ADDA system.
How big an image do you normally acquire? In an earlier reply I talked about the limitations of printers for the number of pixels. Another issue would be the amount of noise in any given pixel for the amount of time dwelt there. I fear that if the resolution is pushed too far that the noise in the pixels would be so high that you would need to average them with their neighbors to get the noise down, and then you might have well scanned fewer pixels for a longer time to begin with.
Comments?
At 11:07 AM 10/29/1999 -0500, you wrote:
} Many thanks to all for numerous replays to my question! } Unfortunately there is no simple way to print... } So, for now I will just use screen capture, and it is not } a good way to use our Fujix Pictrography 3000 printer with } 3800*2759 print pixels. } I just hope that PGT will be more user friendly in future and } release file conversion utility (to do it by myself will be too } time consuming). } } Vladimir
Cambridge Healthtech Institutes Fourth Annual Advances in MOLECULAR LABELS, SIGNALING & DETECTION: Enhancing Sensitivity, Accuracy, and Speed June 12-13, 2000 The Capital Hilton Washington DC
Extending the limits of assay sensitivity and accuracy, while also meeting the demand for greater throughput and/or lower cost, requires tightening engineering specifications, but also acquiring innovative techniques and systems. The development of new probes and labels, homogeneous assay designs, and approaches which allow for the direct detection of compounds or specific binding events are having an impact in basic research, diagnostic and drug development segments. Novel fluorescent and luminescent technology are also critical for the implementation of greater speed and automation. Advances in miniaturization, including microarrays, are having a significant impact in meeting these goals. These advances are being applied to the detection, quantification and localization of gene sequences, proteins, infectious organisms, contaminants and a variety of other targets.
Researchers are encouraged to submit a proposal for presentation. Recommendations for other speakers to be considered are also welcomed. Potential topics include novel developments and new methods for:
Instrumentation Biosensors Electrochemical Sensors Fluorescent and Luminescent Assay Systems Spectroscopy
Labels and Probes New probes Novel labels Quantum dots
Detection Direct (non-amplified) quantitation Optical sensing and detection Single molecule detection Ultrasensitive detection Ultrafast DNA detection
Applications Clinical molecular diagnostics High throughput genotyping High throughput drug screening Homogeneous assays
Please submit proposal or suggestions by e-mail or fax to:
Mary Chitty Conference Director e-mail: mchitty-at-healthtech.com fax: 617-630-1325
For full consideration, please submit proposal or suggestions by November 19, 1999.
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Dear Linda As far as I know, the original supplier of kainic acid lost the algae this compound was obtained from and thus the extreme shortage. Sigma at one time about 6 months ago was offering kainic acid at very high prices and only had 5mg in stock. I don't know of any company to purchase it from. We have looked but found nothing also. If your friend has some very generous colleagues they may persuade them for some. That's what we did. I also have not found an alternative to the compound. I have heard a rumour that over 100uM ampa (s) zwitterion will activate kainate receptors, but I don't know if this is true. If you find anything helpful, I would appreciate knowing the answer to your question for sources of kainate or even substitutes. Sincerely, Linda Chicoine Cognetix, Inc.
id AV23430; Sun, 31 Oct 1999 12:30:36 +0200 To: harrison-at-math.uio.no Message-Id: {1981746242987316.110491847413-at-math.uio.no}
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Bill - Its not easy, but you knew that. Some virus have characteristic shapes (e.g. bullet shapes) and they cannot be confused with cell structures. The most difficult are the numerous, small isometric virus. If they appear in the nucleus of the cell its a give-away, or if they are repeatably observed passing through cell walls. If those "virus" grow in good numbers, plant tissues can be stressed (part drying before fixation) and virus may then form a crystal array. I expect that immune tagging techniques could be used, but I have little experience with those. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, October 30, 1999 5:08 AM, William McManus [SMTP:billemac-at-biology.usu.edu] wrote: } } } To all listers: } } We have a very limited amount of protozoan sample which appear to have } virus-like particles within the cells, but we need to know more conclusively } if the particles are virus before we continue. Does anyone know of a method } using EM to identify virus in tissue? We presently have material available } and could run either a pre or post embedding technique. } } William McManus } Supervisor } Electron Microscopy Facility } Department of Biology } Utah State University } Logan UT 84322-5305 } } billEMac-at-cc.usu.edu } 435-797-1920 }
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