Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 43-22 85 61 89 (work) + 43-22 43 83 23 (privat) Fax.: + 43-22 85 47 26 e-mail.: abrech-at-bio.uio.no
Comments: Authenticated sender is {wesleysm-at-biology.und.ac.za}
Dear Fred
On the cryo side I recommend Patrick Echlin's James Wesley-Smith Electron Microscope Unit George Campbell Building University of Natal Durban, South Africa
Comments: Authenticated sender is {wesleysm-at-biology.und.ac.za}
Ooops. Let's try again! Dear Fred On the cyo side I recommend Patick Echlin's Low Temperature Microscopy and Analysis (Plenum Press) as an "all rounder" reference textbook on the subject, including a chapter on cryomicrotomy.
Wish you and membres of this listeserver a prosperous and aberration-free 1996.
James Wesley-Smith Electron Microscope Unit George Campbell Building University of Natal Durban, South Africa
I am trying to sell our TEM. I have advertised in Microscopy Today and Journal of the Microscopies and have had some interest, but I have not gotten any bids. Our scope is five years old and in great condition. I have heard that there is a flood of TEMs on the market, so the value of TEMs has dropped. I would be interested in talking to anyone who has sold or bought a scope resently. Duke has told us that we should take bids. Would it be better for us if we set a price we could advertise?
Happy New Year to Everybody! - Does anyone have a reference for karyotyping rat cells? Any reference would be welcome! Thanks in advance. Peter P. Molnar, M.D., Ph.D. molnarp-at-lib.dote.hu
For the past few months we have been archiving information from this mail server, and other sources, that related to biological microscopy. There are now 45 topics available. We hope that others can make good use of this information.
go to: http://www.biotech.ufl.edu/~emcl. Then click the Wizard on the Tips and Tricks block.
We will include any contributions that you might want to make and will appreciate any comments that you might have. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
Recently there was a thread about both commercial and home-made Polaroid negative clearing tanks/racks. I apparently didn't keep a copy of the thread. Could someone point me to the listserver archives or email me a copy of the thread?
Thanks,
Henk Colijn
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- An optimist believes that we live in the best of all possible worlds. A pessimist fears this is true.
Although not strictly a microscopy question, perhaps someone has an answer for me. We are considering buying a scanner for a Macintosh computer. I have a catalog that lists them from $300 to $3000. Obviously, there must be differences. Has anyone had any experience with scanners (Relisys, Umax, Epson, Microtek)? Color vs B&W? Scan speed? DPI? How about software? What's a good OCR program? Etc?
Bob
Robert R. Wise, PhD Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
For several years now three SEM labs here at the Univ of Michigan have not been using sulfite baths for treating Poloroid P/N negatives. Instead, they merely rinse them for an hour or so in warm running water (only a slow flow is needed-just enough to keep the water lukish) and then hang them up by the corner to drain and dry on spring-clip-type clothespins that are strung on a piece of rope or wire. This eliminates the cost of the sulfite bath, the problems of disposing of the spent bath liquor, and the ungodly mess that students always produce by splashing the sulfite solution all over the lab. Try it, you might find it satisfactory for your purposes. Wil Bigelow (bigelow-at-umich.edu)
Can anyone provide a few micrographs showing the gross presence/distribution/morphology of mitochondria in the human hair shaft? For teaching and demonstration purposes only.
I am studying the microfractographic network and the mineral content of bricks. In doing so it is my hope to use confocal microscopy to analyse the microfractographical network of the bricks. However, I am not certain of the common techniques used to identify the minerals that make up the bricks and how to associate different mineral grains to the fissures that may traverse them. It is my hope that I can develope a multi-digital image analysis of the bricks' fracture network and its mineral composition. For mineral identification I have found a number of techniques discussed in the literature such as polarised light microscopy, acoustic microscopy, SEM (Backscattered electron imaging and energy diepersive X-Ray mapping) and uranium -induced fission tracked micro-mapping. I would like to request any information on these techniques or any such others that may be used in conjunction with CLSM for mineral identification of the composition of clay bricks as to form a multi-image analysis technique.
Thank-you,
Felicity Lawrence e-mail : f.lawrence-at-qut.edu.au
} To all, } } Although not strictly a microscopy question, perhaps someone has an } answer for me. We are considering buying a scanner for a Macintosh } computer. I have a catalog that lists them from $300 to $3000. Obviously, } there must be differences. Has anyone had any experience with scanners } (Relisys, Umax, Epson, Microtek)? Color vs B&W? Scan speed? DPI? How } about software? What's a good OCR program? Etc? } } Bob } } } Robert R. Wise, PhD } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-vaxa.cis.uwosh.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } We are running a Umax Power-look with a transparency scanner for doing negatives. It come with all the right software including Adobe Photoshop for either PC or MAC. I think it was about $2600. directly from Umax. We have been very happy with it so far. Another lab has the same scanner running on an SGI Indy. Apparently the software is more tricky on that platform but the results have been very good. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
I am using a Hewlett Packard ScanJett IIc, about 800 dpi with some interpolation, color / bw. This is an "older" model, so I bought a used one for 500 sFr. (swiss franks).
- OCR even for small printed text (as in journals) is satisfying with OmniPage. - Microscopic slides *can* be scanned, resulting in about a 30 fold magnification.
Personally I am very satisfied with this device. Good Luck, Wolf.
} To all, } } Although not strictly a microscopy question, perhaps someone has an } answer for me. We are considering buying a scanner for a Macintosh } computer. I have a catalog that lists them from $300 to $3000. Obviously, } there must be differences. Has anyone had any experience with scanners } (Relisys, Umax, Epson, Microtek)? Color vs B&W? Scan speed? DPI? How } about software? What's a good OCR program? Etc? } } Bob } } } Robert R. Wise, PhD } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-vaxa.cis.uwosh.edu
MSA is seeking nominations for its major annual awards, to be presented at the 1996 Annual Meeting in Minneapolis, MN in August. Nominations must be received by January 15, 1996, and should consist of a brief description of why the nominee is deserving of the award and, if possible, a CV or resume. Additional notes of support from colleagues are also helpful.
The major MSA awards are as follows:
MSA Distinguished Scientist Awards (Physical and Biological): given for unique and distinguished contributions to microscopy, imaging, and/or compositional analysis.
Burton Medal: given for the most important contribution(s) in the fields of microscopy, imaging, and/or compositional analysis during the last five years by a person under the age of 35 on June 30, 1996. Preference will be given for work carried out in North America.
Morton D Maser Distinguished Service Award: given to honor significant volunteer service to MSA over a period of years and generally in more than one area.
Outstanding Technologist Award: given to honor outstanding contributions by a technologist.
Nominations or questions can be addressed to:
Ernie Hall MSA Physical Sciences Director GE CRD, PO Box 8 (1 Research Circle), Schenectady, NY 12301 e-mail: hallel-at-crd.ge.com fax: 518-387-6972 phone: 518-387-6677
Conly Rieder MSA Biological Sciences Director Wadsworth Center, Emipre State Plaza, PO Box 509, Albany, NY 12201 e-mail: rieder-at-wadsworth.org fax: 518-486-4901 phone: 518-474-6774
The Ocular Cell Transplantation Laboratory in the Department of Ophthalmology at UMDNJ, New Jersey Medical School has the following job openings:
ELECTRON MICROSCOPIST: The individual will be expected to perform all aspects of TEM from harvesting animal tissue to preparation of publication prints. In addition, some specialized EM techniques will be performed such as immunocytochemistry and in situ hybridization. Animal work is required and will involve assisting in surgeries and euthanasia. We are seeking a skilled electron microscopist with a minimum of three years experience after receiving a bachelor's degree.
RESEARCH TEACHING SPECIALIST: This individual will be expected to harvest tissue and prepare it for light and electron microscopy. Embedding media will mainly involve epon, JB4, or lowicryl. Animal work is required. A bachelor's degree plus one year of relevant experience after receipt of the degree are the minimum requirements for this position. Individual must have experience embedding and sectioning tissue embedded in epon and JB4. Experience with black and white photography highly desirable.
MICROSCOPIST: Medjet, Inc., a biotech company specializing in the development of ophthalmic instruments is seeking a skilled microscopist to conduct animal studies. This position requires an individual with skills in both TEM and SEM. The individual will be expected to work independently and prepare and present data at weekly meetings. The position requires commuting between UMDNJ in Newark to the Medjet facilities in Edison, NJ (about an hour drive). A PHD or MA is required with a minimum of 3 years relevant experience.
Interested individuals should mail or fax their resumes to:--
Does anyone have a reference to a description of the actual method of making coverslips? Is it true that they are float glass? Wouldn't that mean that one side might be different from another (not optically, but maybe chemically)? Inquiring minds need to know...
Bob, we are using extensively flatbed scanners for digital imaging of our negatives (fluorescence and histology color LM, TEM and old Polaroids from my FSEM). My suggestion: Buy an Agfa Arcus II for true 8-12 bit graytone or 24-48 bit color "output" in TIFF. AGFA is experienced, has excellent color handling and will be around for a long time for support. The street price of US$ 2,000 (any US catalog company, i.e., The Mac/PC Zone 1-800-248-0800) includes a "full version" of Photoshop 3.04 which reads and writes 8-12 bit Tif files. TIFF is the MSA standard. Both versions (PC and Mac) of Photoshop 3.04 are now available and allow us to work with 12-bit Tiff's (at my microscope on a PC and at my networked office with a Mac). The scanner comes with AGFA's FOTOLOOK acquisition software which is opened from within Photoshop: Files-Acquisition.
Please consider: Not spatial resolution but "contrast resolution" is important for the acquisition of digital image data, i.e., if you read more intensity steps you can work with more intensity levels per pixels and can "see" smaller contrasts. Our experience is that all microscopes can deliver 12-14 bit contrast resolution. The jump from 8-bit to 12-bit gives you an increase in contrast resolution of 1:16 which is adequate for utilizing the full range of contrasts found in (TEM, Polaroid) negatives. Their is already some software out for the handling of 12-bit image data and I believe that soon many other software packages will available for working at the level of 12-bit contrast resolution already available in many microscopes, e.g. AFM microscopes, good CCD cameras for TEM and several SEM acquisition systems. Even, if you cannot handle 12-bit data at this time and may have to wait for an upgrade of NIH image, a foreseeing investment in adequate hardware will pay of very soon when 12-bit contrast resolution will have a common place in microscopy imaging.
Optimal scanning of negatives requires that the negatives are exposed 1-2 stops more than for ordinary photographic printing (high lights should be "covered" and not be empty, dark areas should be very dark. Good scanners can handle more than 3.0 optical density which is "very dense" for a photographic negative). Also the scanning procedures must be adapted in order to take advantage of the full contrast transfer function of the negatives as well as the scanner. I am in the process of putting a report together at my WWW site on our experience with 8-bit versus 12-bit negative scanning.
I am looking for a protocol for the embedding of yeast for TEM. I made a few attempts and the results have been mixed. The biggest problem is that the cut sections look like Swiss cheese. Where yeast should be there are holes. The fixation appears to be fine in the yeast that do not fall out. I am treating the cells with KMnO4. Over weekend infiltrations with Spurr's or Epon 812 have not worked. Any suggestions would be helpful.
} We are running a Umax Power-look with a transparency scanner for doing } negatives. It come with all the right software including Adobe Photoshop } for either PC or MAC. I think it was about $2600. directly from Umax. We } have been very happy with it so far. Another lab has the same scanner } running on an SGI Indy. Apparently the software is more tricky on that } platform but the results have been very good.
Dear Greg, What is the resolution of this scanner? How accurate is the quan- titation of the OD's? Would it be a suitable substitute for the Perkin- Elmer flatbed microdensitometer? TIA. Yours, Bill Tivol
If black and white prints are to be scanned, the Apple one Scanner (up to 1200 dpi) cost now less than 700 dollars and is very good with OFOTO software. Equivalent scanners (e.g. LaCie are just a good), but I am not sure about PC equivalent and would certainly like to know the outcome of recommendations. Images in the page below were scanned with the Apple one scanner at 300dpifrom black and white prints without any manipulation (filtering, etc).
****************************************************************** *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * * {Fermin-at-tmc.tulane.edu} Prof. Pathology & Otolaryngology * * http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html * ******************************************************************
} I am looking for a protocol for the embedding of yeast for } TEM. I made a few attempts and the results have been } mixed. The biggest problem is that the cut sections look } like Swiss cheese. Where yeast should be there are holes. } The fixation appears to be fine in the yeast that do not } fall out. I am treating the cells with KMnO4. } Over weekend infiltrations with Spurr's or Epon 812 have } not worked. Any suggestions would be helpful. } } Thanks, Greg Rudomen } Greg-at-umic.umic.sunysb.edu
RESPONSE:
Hi Greg, I've worked with Candida albicans and Saccharomyces over the years and rarely had problems using KMnO4 fixation and embedding in either Spurr's or Epon-type resins. Check that the cells are completely dehydrated (is the alcohol or acetone really absolute?) and that there is no water in the resins.
I have used the following protocol: After fixation, rinse 3x in distilled water 20 min ea. If individual cells, suspend the cells in 2% agarose at 45 C, pour onto glass slide and allow to solidify. Cut into small cubes. Suspend briefly in dist water and then dehydrate in ethanol series (25 min ea), 25, 50, 75, 95, ABS x 3 changes. Prop oxide 3 x 25 min. I use a l:l mix of Epon/Spurr's resin made up for long pot life. Infiltrate as follows: 2 PO/1 resin for 1 day, equal parts PO/resin for 1 day. Pure resin in capped container at RT 3 changes at 1 day each. Transfer specimens into capsules and polymerize at 60 C for 48 hr.
CALL IF YOU NEED MORE INFO.
############################################################################# John J. Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
I am looking for an iris diaphragm for a Nikon SMZ-10 Stereo microscope. They have discontinued this item, it is a double diaphragm wafer that goes in the microscope stack. The Part number is 76270. I am also looking for a fine focus attachment for the Nikon camera UFX. It fits over the eyepiece of the camera. Larry Albright 419 Sunset Avenue Venice, California 90291 Phone 310-399-0865...Fax 310-392-9222 or albrite-at-netcom.com
Comments: Authenticated sender is {wesleysm-at-biology.und.ac.za}
Dear Greg You may also want to try freeze-substitution. Although it sounds like more than you bargained for, it works superbly for yeast (personal experience). Because of their small size, they freeze and substitute well, and the preservation attained is worth the effort.
Try and get hold of the article by Baba and Osumi in the Journal of Electron Microscopy Technique, March 1987, Vol 5, number 3, p249.
Good luck.
James Wesley-Smith Electron Microscope Unit George Campbell Building University of Natal Durban, South Africa
One user's opinions: 1. Consider the media you will be scanning. Some scanners do better at transparencies than others (nikon has a slide scanner for 3K$), some are designed for large format (Umax has a 17 x 12 inch for 7k$), some do black and white on the cheap (HP has one B&W for $400).
2. Consider the destination of the image. There is no use buying a 30 bit high definition scanner if all you will do is newsprint. A 300dpi 256 gray scanner goes a long way with laser printers and costs $400.
3. Consider the resolution you need. If you are going to scan stamps that are 1 inch, and display them at poster size, you need a very high res scanner (1200 dpi or more) if you are going to scan 8 x 10 positives and display them in a small window, then 600 is plenty, 300 adequate.
4. Consider the speed. Some scanners take 30 seconds to scan one page. This can be a real pain if you have any number of images to scan. Some are available with a document feeder, which is a nice thing to have, but not if all your scans will be of pictures (which must be hand fed anyway) "Single pass" scanning means that all 3 colors (red green blue) are captured simultaneously--this is faster (HP scanjet 4 offers this)
5. Real Res. Most scanners interpolate the dpi at high res. This is ok, but non-interpolated scans are better. Be willing to pay more for a hardware 600dpi than a software 600dpi.
Does anyone have any experience or advice on substrates suitable for ultramicrotomy of settled organisms for TEM?
We have someone culturing small marine sea-squirts (Ascideans) who wants to cut ultrathin sections. These form spreading colonies about 1 mm in height. I have seen references somewhere to Mercox/Mercanox? but do not know its source/supplier. Also can this be cut on glass knives - marine organisms are notorious harbourers of small sand particles which do not go well with diamond knives!
Any comments welcome.
Keith Ryan Plymouth Marine Laboratory Citadel Hill Plymouth PL1 2PB, UK
I am prepared to purchase a microtome for LM. I have info on the Zeiss HM 315 and Zeiss HM 325 (the one with specimen retraction. I also have info on the Leica Histocut 820. I wish to section chick embryos from very early stages, in paraffin blocks for immunostaining purposes. I have been using a very old AO Spencer. I am interested in "bang for the buck" value. Is the specimen retraction feature worthwhile for paraffin block sectioning? Are the disposable blades as stable as the stainless steel knives or can one get sectioning artifacts when using very small block faces? Any advice or suggestions? Darrell Wiens University of Northern Iowa
} I am looking for a protocol for the embedding of yeast for } TEM. I made a few attempts and the results have been } mixed. The biggest problem is that the cut sections look } like Swiss cheese. Where yeast should be there are holes. } The fixation appears to be fine in the yeast that do not } fall out. I am treating the cells with KMnO4. } Over weekend infiltrations with Spurr's or Epon 812 have } not worked. Any suggestions would be helpful. } } Thanks, Greg Rudomen } Greg-at-umic.umic.sunysb.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
Greg, See: Anderson et al., 1991 J EM Tech. 18:172 for a protocol using periodate treatment of the wall.
and, Byers & Goetsch, 1991 Methods in Enzymology 194:602 for a method using enzymatic treatment of the wall. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
} Optimal scanning of negatives requires that the negatives are exposed 1-2 } stops more than for ordinary photographic printing (high lights should be } "covered" and not be empty, dark areas should be very dark. Good scanners } can handle more than 3.0 optical density which is "very dense" for a } photographic negative). Also the scanning procedures must be adapted in } order to take advantage of the full contrast transfer function of the } negatives as well as the scanner. I am in the process of putting a report } together at my WWW site on our experience with 8-bit versus 12-bit negative } scanning. } } Best regards Klaus } } ****************************************************************************** } * : * } * Klaus-Ruediger Peters, Ph.D. : WWW Home Page: * } * Director, Molecular Imaging Laboratgory : * } * Biomolecular Structure Analysis Center : Molecular Imaging Laboratory * } * University of Connecticut Health Center : http://panda.uchc.edu/ * } * 263 Farmington Ave. : htklaus/index.html * } * Farmington, CT 06030-2017; U.S.A : Differential Hysteresis * } * : Processing Demo at http:// * } * Tel: (203) 679-3977; Fax: (203) 679-1989 : panda.uchc.edu./htbit/indiv/ * } * e-mail: Peters-at-BSAC.UCHC.EDU : /software_docs/dhp.html * } * : * } **************************************************************************** } ** } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } we have partially overcome this problem by overlaying negatives with photographic contrast filters or neutral density filters. This may not be appropriate for very high res. work but is no problem for video display and printing to a laser printer -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
Our department has about a dozen old Spencer light microscopes--monoccular, black laquer with lots of brass, probably pre-WWII. Does anyone know if there is a market for such things out there? Are there 'antique' microscope dealers who might be interested in these? They are not very functional but too nice to throw away.
Bob
Robert R. Wise, PhD Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
In response to Klaus Peter's eloquent plea for 12-bit versus 8-bit scanners:
While I would not want to be thought of against precision, it must be admitted that there are some limitations to using 12-bit data, namely greater storage (and display?) requirements. So one should consider this step carefully.
Assuming that it is done correctly, 12-bit data is digitized to 1 part in 4,000 while 8-bit data is digitized to only 1 part in 256. Therefore, to make proper use of such precision one must have data of similar precision. Although many considerations can reduce the precision of recorded data: measurement noise, low contrast, it can never be better than the statistical limitations placed by quantum mechanics on the number of events counted (grains of silver (assuming that they are all of the same size which whey aren't)/pixel or photons/pixel or electrons/pixel). Clearly, this all depends on the size of a pixel: larger pixels imply the chance to count more quanta (but they also mean lower scanner resolution) so you may need less intensity resolution if you have greater real (non interpolated) spatial resolution.
Now it is clear that, for instance, in STM, where a 0.05 nm height resolution can be detected and the piezo may have a height sensing range of many microns, there is no difficulty in obtaining 12-bit data (or even 16-bit data) although you may have trouble displaying all of this "depth" to a human observer at one time. Likewise, CCD sensors having noise levels of +/- 3-4 RMS electrons/pixel and full-well signal levels of 300,000 electrons/pixel easily satisfy the 4,000:1 (more like 100,000:1 possible) requirement as long as you use a long enough exposure to actually record at least 20,000 electrons/pixel. However, in confocal microscopy, 256 photons/pixel is often quite a high signal and digitizing from a color slide of a fluorescent image recorded on high speed film (depending on original magnification and pixel size. Remember, 1200 DPI implies 20 micron pixels.) one may find it difficult to find even 10 distinguishable levels.
So far we have only spoken of linear digitization (as is appropriate to scanners which usually make every effort to be linear). However, for signals degraded mainly by statistical noise and recorded by the direct digitization of electron signals, the separation between "meaningful" grey levels (those separated in intensity from neighboring levels by an amount at least equal to their standard deviation?) the difference between the first two such levels (1 event/pixel and 4 events/pixel) is 3 event/pixel whereas that between the 15th level and the 16th is 31 events/pixel. In other words, if we are only interested in recording the INFORMATION in a signal limited only by quantum noise (SEM? Confocal?), we could first take its square root and then digitize this. This could be done using only one half of the number of bits that would have been necessary to preserve the information in a linear signal and each SQRTBit would be a "real" grey level.
All this said, one must admit that getting real 8-bit data requires more than the use of an 8-bit DAC (0.4% illumination stability, low-noise detector, freedom from digital electronic interference, proper bandwidth for sampling rate in both the electron and optical parts of the information path, etc) and many scanners may not provide the performance they claim. This might explain much of the difference claimed to exist between 8-bit and 12-bit results.
(To any owners who have read this far: have you any MEASUREMENTS on scanner performance on known images?)
SEM might provide a good case in point: The SE signal variation associated with small features is often less than 1% of the total signal. As "seeing" a small feature with only 1% contrast requires collecting at least 1/0.01 x0.01 = 10,000 electrons/pixel, it might seem worthwhile to use 12-bits to preserve these small variations. However, unless there are "holes" in your specimen from which virtually no signal is obtained (i.e. if your signal does not have "important" high contrast), little would be lost if one digitized only the 256 levels nearest in intensity to those of the small feature and a positive improvement would be gained (all the information that could possibly be coded in up to 65,000 detected electrons vs. 10,000) by digitizing the square-root of the signal into only 8 bits. In either case you would have to "process" the signal before you displayed it on the screen.
Just some thoughts. Sorry that they were so long but I felt a trend to Digital-One-up-man-ship...
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
Dear Colleages, I'm having trouble locating a simple item - those plastic ball and stick models for making models of simple crystals. I need some models for a course I'm teaching in the spring, and so far the models I've seen in the scientific/lab products catalogs I've looked in are a bit disapointing. By that, I mean they are more oriented toward molecular models, and the emphasis in the models are on the bonds, not the arrangement of the atoms, which is what I'd rather stress for this class (intro to semiconductors). Does anyone out there know of any good sources for ball and stick models? Thanks
} } I am looking for a protocol for the embedding of yeast for } TEM. I made a few attempts and the results have been } mixed. The biggest problem is that the cut sections look } like Swiss cheese. Where yeast should be there are holes.
Dear Greg, I had the same problem with pollen grains, and it arises from the difference in hardness between the specimen and the resin. Our solution was to experiment with different proportions of the constituents of the resin until its hardness matched that of the specimen. Of course, this has to be a trial-and-error process. Good luck. Yours, Bill Tivol
} } K.-R. Peters wrote: } } } } Optimal scanning of negatives requires that the negatives are exposed 1-2 } } stops more than for ordinary photographic printing (high lights should be } } "covered" and not be empty, dark areas should be very dark. Good scanners
} Greg Erdos responded: } } we have partially overcome this problem by overlaying negatives with } photographic contrast filters or neutral density filters. This may not be } appropriate for very high res. work but is no problem for video display and } printing to a laser printer } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- } Greg Erdos Phone: 352-392-1295 } Scientific Director, } ICBR Electron Microscopy Core Lab } 218 Carr Hall Fax: 352-846-0251 } University of Florida E-mail: gwe-at-biotech.ufl.edu } Gainesville, FL 32611 } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
Thanks Greg, you make an important point concerning acquisition and display:
The application of contrast or neutral filters in photography shifts the brightness (level) or contrast (slope) of the image data to a range at which the eye is more perceptive (details in high lights may be preserved, but reduced in the shadows or visa versa, or both may be compressed). These procedures do not produce more "image detail" than the negative contains but modifies the display of the details. If during image acquisition (negative exposure) the highlights in the negatives were underexposed (or the shadows overexposed) then the possibly lost detail information can not be recovered, neither analog nor digitally.
It is therefore important (for both analog as well as digital imaging of the negative) to the set the brightness level of the negative so that it captures as much information as possible. Generally, the contrast transfer function (slope) is not changed, i.e., through variation of developing conditions or change of film/developing combination. But why don't we normally care much about optimal exposures (or acquisition)?
Photography (analog) had long accepted that there is more information in a negative (raw data) than the eye can "see" ("image" being that part of the displayed image data which the eye can perceive and recognize). It developed many useful tools for enhancing the contrast of certain image components of the negative so that they become visible, but consider our following findings. Measurments of the intensity ranges of contrast patterns in digitized EM negatives and the contrast ranges required for visual display of these contrast patterns (as images) indicates that an optimally exposed TEM negative has an information depth of approximately 12 bit, or a 12-bit contrast resolution whereas the "image" has merely an approximate 6-bit contrast resolution. We find in digital image data from nearly all microscopies that the detail contrasts (structures of lowest contrast levels) occupy only 1-10 % of the overall contrast range of the image data whereas we need about 25% of the contrast range of an image for visual recognition (not perception) of a contrast pattern (in an amorphous environment). This means, most of the low-contrast data components (which constitute the high-resolution details) are not visible in a conventional image of the data. Unfortunately, we have adapted to this fact and tend to acquire images at an insufficiently low contrast resolution level equal to that of our visual system because we are not missing anything in the "empty" high-lights or empty shadows although our microscopes can acquire images at much higher contrast resolution. On the other hand, these facts underline why digital acquisition (from the imaging sensor or from the negative) should be done at a 12-bit level instead of an conventional 8-bit level. Many manufactures of acquisition and processing equipment start to provide a 12-bit "output" level, 12-bit scanners are now offered at catalog firms, and Photoshop is now available for PCs and Macs with 12-bit TIFF capabilities (version 3.04).
The basic idea of scanning negatives is to "preserve" all information of the negative with a linear transfer function, irrespectively if the eye on the negative or print can see it or not, and then use digital imaging for the display of all the available image information. Thus, when exposing a negative for digital scanning, we should capture the contrasts of interest with the largest possible contrast range. Since good scanners can penetrate the darkest black of a negative (max. OD = 3-3.5) capturing the details in the shadows, we should expose more than conventionally done in order of acquiring also as much detail as possible in the high-lights.
Additionally, one should consider that laser printing (HP LaserJet 4 providing 5-bit gray levels and 100 lines/inch) combined with the LazarPrint expansion can print 8-bit gray levels at 300 lines/inch, thus provides a good output for detail rich images at the 1Kx1K level (I tested recently several printers and reported the results in my home page).
I got a number of good responses on my question regarding flatbed scanners. Everybody seemed pleased with the particular one they owned--there were no clear favorites. A couple of tips: 1) Make sure your computer has the right output. All of the scanners I have looked at have a SCSI 2 output while my 2.5-year-old Mac only has a SCSI. I have been told that I have to upgrade my computer in order to even hook it up to a SCSI 2 scanner. I'm still investigating this. 2) Make sure you get the software you need to manipulate the scanned image or text. A lot of scanners come with software bundles. 3) Look into warranties. Some devices have none--others are up to 2 years (longer?). 4) Photoshop comes in two versions--LE and 3.1. The latter is more powerful (and expensive) than the former. 5) Transparency adapters and document feeders are usually extra. Both can be useful.--if you need them. 6) Read the first response (from {sco.umc2-at-Mail.health.ufl.edu} ). It contains a lot of good basic advice. 7) Good luck.
Bob ________________
One user's opinions:
1. Consider the media you will be scanning. Some scanners do better at transparencies than others (nikon has a slide scanner for 3K$), some are designed for large format (Umax has a 17 x 12 inch for 7k$), some do black and white on the cheap (HP has one B&W for $400).
2. Consider the destination of the image. There is no use buying a 30 bit high definition scanner if all you will do is newsprint. A 300dpi 256 gray scanner goes a long way with laser printers and costs $400.
3. Consider the resolution you need. If you are going to scan stamps that are 1 inch, and display them at poster size, you need a very high res scanner (1200 dpi or more) if you are going to scan 8 x 10 positives and display them in a small window, then 600 is plenty, 300 adequate.
4. Consider the speed. Some scanners take 30 seconds to scan one page. This can be a real pain if you have any number of images to scan. Some are available with a document feeder, which is a nice thing to have, but not if all your scans will be of pictures (which must be hand fed anyway) "Single pass" scanning means that all 3 colors (red green blue) are captured simultaneously--this is faster (HP scanjet 4 offers this)
5. Real Res. Most scanners interpolate the dpi at high res. This is ok, but non-interpolated scans are better. Be willing to pay more for a hardware 600dpi than a software 600dpi. __________________
Sender: bob-at-befvax.uchicago.edu
What resolution do you need. Many of the inexpensive scanners have a very small numerical aperature which leads to line broadening. So a step size of (for example) 5 microns will not resolve a 5 micron line in single (or two) pixels but rather 10 or more. The requirement for a large numerical aperature coupled with dimensional stability is what makes high resolution cost so much.
Bob J. ______________
Response from John Bozzola:
We use a Mac IIci attached to a LaCie 600 dpi color scanner (it's equivalent to an Epson 300) to occasionally digitize prints. It cost $1,800 - a similar unit may now be purchased for around $600. I have used it for over 3 years now with no problems whatsoever. 97% of the scanning is done in the bw mode set to no more than 300 dpi. The program most often used to drive the scanner is Adobe PhotoShop. As an OCR program, I use WordScan Plus. It's OK, a bit slow, but it is also 3 years old and a newer version may be better. I rarely use this program. However, when the documents are of high quality (typed characters no smaller than 9 point) then it is a great time saver. There are better OCR programs out there than this one. _________________
Our Graphics Artist uses a Macintosh computer with a Hewlett Packard scanner which he likes very much. The one we have is color and has a 400 optical DPI, although the new Hewlett Packard has a 600 optical DPI. It comes with Deskscan software and a limited version of PhotoShop. If you want to use color, though, you need to have the full version of PhotoShop.
Hope this is helpful.
Kathy Stangenberg Ted Pella, Inc. _______________
I have a AGFA StudioScan IIsi scanner with Transparency module. I think that it is a very good scanner. I using it every day. Some technical data:
- Max. res. 400(H) x 800(V) optical 2400(H) x 2400(H) ppi through interpolation
MAC PC SM QM SM QM Preview colour 16 16 15 15 A4 colour 200 ppi 54 54 32 39 A4 colour 400 ppi 96 104 86 126 15x10 cm colour 400 ppi 44 54 30 38 A4 grey 200 ppi 17 18 12 12 A4 grey 400 ppi 39 42 39 65 A4 line art 400 ppi 20 34 20 34
SM=Speed mode (in seconds) QM=Quality mode (in seconds)
Software:
- OmniPage Direct OCR (I suggest you to buy Recognita OCR software, because it is the best as I think) - FotoTune - PhotoShop LE
The one chosen to be included in the Kodak Imaging Station is the high end EPSON with or without the transparency adapter. I have seen the output from this equipment rated at an interpolated 4800dpi , and it is very impressive. When prints are made on a dye-sub printer (Kodak or Codonics), it is hard to tell that they were not made from a negative in a conventional darkroom.
Regards, Skip ___________________
We are running a Umax Power-look with a transparency scanner for doing negatives. It came with all the right software including Adobe Photoshop for either PC or MAC. I think it was about $2600. directly from Umax. We have been very happy with it so far. Another lab has the same scanner running on an SGI Indy. Apparently the software is more tricky on that platform but the results have been very good.
We have an HP Scanjet 3c/T scanner on a Powermac 9500 used for image analysis and presentation purposes. It is a superb accessory. You can scan virtually anything that is 8.5 X 11 or smaller with it, although it does not substitute for a good slide scanner. Since it has the transparency adapter gels come out very nicely. It has its own miniprogram for image adjustment, but if you are doing serious work we have an Adobe suite (Illustrator, Pagemaker & Photoshop) installed on the Mac and that is what just about everyone uses. For many types of text (but definitely not all) you do not even need OCR. If you can afford it, it seems to be the way to go. I am a user and do not have any financial connection to Hewlett - Packard.
Bruce Cutler, Microscopy Laboratory, Univ. Kansas, Lawrence. _________________
We have experience with some HP-IIcx's. One is on a Mac, the other on a IBM PS/1. Both seem to work quite well. They ran about $1000 over a year ago. I think they support 1200 dpi.
They came with minimal software. The software allowed scanning images well enough. But optical character recognition required third-party software. That may all have changed.
Warren E. Straszheim E-Mail: wes-at-ameslab.gov (or: wesaia-at-iastate.edu) ___________________
I am using a Hewlett Packard ScanJett IIc, about 800 dpi with some interpolation, color / bw. This is an "older" model, so I bought a used one for 500 sFr. (swiss franks).
- OCR even for small printed text (as in journals) is satisfying with OmniPage. - Microscopic slides *can* be scanned, resulting in about a 30 fold magnification.
Personally I am very satisfied with this device. Good Luck, Wolf. ______________
We use a UMAX 1260 color flatbed scanner, which will scan to 3000 dpi, and we love it. It is a three pass color scanner, but the speed isn't slow enough to justify another $1000 for a one pass scanner. It works as a plugin to Photoshop.
Also, we use OmniPage Pro for OCR, and again, it's great!
Any questions, just email me.
Scott ________________
Bob, we are using extensively flatbed scanners for digital imaging of our negatives (fluorescence and histology color LM, TEM and old Polaroids from my FSEM). My suggestion: Buy an Agfa Arcus II for true 8-12 bit graytone or 24-48 bit color "output" in TIFF. AGFA is experienced, has excellent color handling and will be around for a long time for support. The street price of US$ 2,000 (any US catalog company, i.e., The Mac/PC Zone 1-800-248-0800) includes a "full version" of Photoshop 3.04 which reads and writes 8-12 bit Tif files. TIFF is the MSA standard. Both versions (PC and Mac) of Photoshop 3.04 are now available and allow us to work with 12-bit Tiff's (at my microscope on a PC and at my networked office with a Mac). The scanner comes with AGFA's FOTOLOOK acquisition software which is opened from within Photoshop: Files-Acquisition.
Please consider: Not spatial resolution but "contrast resolution" is important for the acquisition of digital image data, i.e., if you read more intensity steps you can work with more intensity levels per pixels and can "see" smaller contrasts. Our experience is that all microscopes can deliver 12-14 bit contrast resolution. The jump from 8-bit to 12-bit gives you an increase in contrast resolution of 1:16 which is adequate for utilizing the full range of contrasts found in (TEM, Polaroid) negatives. Their is already some software out for the handling of 12-bit image data and I believe that soon many other software packages will available for working at the level of 12-bit contrast resolution already available in many microscopes, e.g. AFM microscopes, good CCD cameras for TEM and several SEM acquisition systems. Even, if you cannot handle 12-bit data at this time and may have to wait for an upgrade of NIH image, a foreseeing investment in adequate hardware will pay of very soon when 12-bit contrast resolution will have a common place in microscopy imaging.
Optimal scanning of negatives requires that the negatives are exposed 1-2 stops more than for ordinary photographic printing (high lights should be "covered" and not be empty, dark areas should be very dark. Good scanners can handle more than 3.0 optical density which is "very dense" for a photographic negative). Also the scanning procedures must be adapted in order to take advantage of the full contrast transfer function of the negatives as well as the scanner. I am in the process of putting a report together at my WWW site on our experience with 8-bit versus 12-bit negative scanning. _____________
If black and white prints are to be scanned, the Apple one Scanner (up to 1200 dpi) cost now less than 700 dollars and is very good with OFOTO software. Equivalent scanners (e.g. LaCie are just a good), but I am not sure about PC equivalent and would certainly like to know the outcome of recommendations. Images in the page below were scanned with the Apple one scanner at 300dpifrom black and white prints without any manipulation (filtering, etc).
KMnO4 fixation? This is interesting, why are you using KMnO4?
Back in the early 1980's we did a pretty frightening study on KMnO4 fixation in fungi (T. M. Hammill, et al. refs available), in which it was found that at concentrations of upto 8.5% KMnO4 the test fungi (Mucor mucedo) continued to grow! (Albeit, abnormally, but ransfer to fresh media resulted in a return to normal growth). So if these fungi continue to GROW at 8.5% KMnO4 how good of a "fixative" is 1-3%?
Additionally, Greg, I agree with Bill Tivol's suggestion that the "swisscheese" effect is due to the fact your resin (1) is too soft - the chitin cellwalls of fungi are hard, and (2) you are seeing poor infiltration of your resin - try a lower viscosity resin, I've had excellent results with fungi using Spurr's Hard or Quetol 651/MNA (Kushida, 1975).
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
I would like to add a couple of comments to the recent thread about negative scanning.
Several people have mentioned using moderately priced flat-bed scanners with transparency adapters. My evaluation of one of these (Epson Scantastic) was that it was useful for quick prints of light (density 1 or less) negatives to show qualitative microstructure. This unit had a non linear response to density, especially over 1. This made the output unsuitable for quantitative analysis and often resulted in "posterization", where large areas with visible contrast differences were all mapped to the same gray level. Check for this by scanning a density step tablet. Scanning periodic features in the negative (i.e. lattice images) resulted in moire patterns (check the FFT of some of your images).
As a result, we chose not to purchase this scanner and added a new PC interface to our old Optronics P1000 rotating drum scanner. With the addition of a new module for the log amplifier that permits a wider range of gain and DC offsets, we can select linear ranges of density of 0.5, 1.0, 2.0, 3.0, and 4.0 with a variable offset of up to 1.5 density units. (This all was supplied by CSI, a small company in England). This provides the ability to get the most significant 8 bits of data per pixel and is free from interference effects.
For the highest resolution, we send the negatives to another lab which has a Perkin Elmer flatbed microdensitometer. These scans are very slow, but have the potential of very high resolution (we have scanned down to 5 microns t0o be certain we oversampled the film).
Best Regards, John
John R. Minter, Ph. D. Phone: (716) 722-3407 Eastman Kodak Company FAX: (716) 477-3029 Analytical Technology Division email: minter-at-kodak.com Rochester, NY 14562-3712
A colleague needs to accurately determine the water content of very small tissue samples (fish embryos at various stages of development). What would be the best way to do this? Thanks for any suggestions. Grace
Registered-mail-reply-requested-by: Ronald.Cohn-at-SYNTEX.COM MR-Received: by mta RVAX.MUAS; Relayed; Fri, 05 Jan 1996 10:05:53 -0800 MR-Received: by mta MEDEC; Relayed; Fri, 05 Jan 1996 10:05:54 -0800 MR-Received: by mta CHUB1; Relayed; Fri, 05 Jan 1996 10:03:19 -0800 Disclose-recipients: prohibited
Microscopists,
Does anyone on the list have tried-and-true methods for eliminating or minimizing background while immunolabeling paraffin-embedded lung tissue? My attempts to label a protein in rat lung using the Vector avidin-biotin alkaline phosphatase kit and the Vector Red substrate development kit have resulted in what I suspect is specific labeling, but also unacceptably high background. Control experiments in which 1) primary antibody has been omitted, 2) spurious avidin binding has been blocked by use of an avidin/biotin blocking kit, and 3) the potential presence of endogenous alkaline phosphatase was blocked by levamisole, have not reduced the background. The background appears to be primarily over connective tissues, and develops simultaneously with specific label.
Any suggestions will be greatly appreciated. If you would respond directly to me, I will compile all reponses and post a summary in a few days. Thanks in advance!
Ron Cohn Roche Bioscience ronald.cohn-at-syntex.com
Subject: Time: 12:09 PM OFFICE MEMO TEM and ID of G+/G- bacteria Date: 1/5/96
To all: Is there a definitive EM method of determining whether an unknown bacterium (in this case a plant pathogen) is G+ or G-? This one gives ambiguous results at the LM level. TIA.
Doug Davis doug_davis-at-maillink.berkeley.edu EML UC Berkeley
} To: William Tivol {tivol-at-wadsworth.org} } From: delannoy-at-welchlink.welch.jhu.edu (Michael Delannoy) } Subject: Re: TEM of yeast } } } } } } } I am looking for a protocol for the embedding of yeast for } } } TEM. I made a few attempts and the results have been } } } mixed. The biggest problem is that the cut sections look } } } like Swiss cheese. Where yeast should be there are holes. } } } } Dear Greg, } } I had the same problem with pollen grains, and it arises from the } } difference in hardness between the specimen and the resin. Our solution } } was to experiment with different proportions of the constituents of the } } resin until its hardness matched that of the specimen. Of course, this has } } to be a trial-and-error process. Good luck. } } Yours, } } Bill Tivol } } } } Greg, } I've got a protocol that works well with yeast (infiltration/embedding): } } 1)Fix in suspension 4%pf/2%ga 3hrs rotating. Spin after ~13K 10 sec. } 2)Rinse in buffer 3 x 10 min (last rinse should be cacodylate) } 3)Postfix 2-4% KMNO4 in caco, on ice 1 hr } 4)3 X 5 min D-H2O } 5)en-bloc 2%UA filtered aq 1hr dark } 6)Quick 50% ETOH then break up pellet into poppy seed size granules } 7)50-70-90% ETOH 5 min ea } 8)3 x 100% ETOH } 9)1:1 Spurrs:ETOH overnight rotating } 10)Vac infiltrate 15 psi fresh Spurrs R.T. 2 hrs } 11)Rotate fresh Spurrs 2 hrs } 12)Repeat step 10 } 13)Change Spurrs and vac infiltrate 15 psi 30 min } 14)Turn on oven cure o\n 60 degrees 15 psi } } You will need an oven that can be pumped down, if not use house vac. } The KMNO4 extracts some ribosomes so that you can see organelles better } than you would with Osmium. Don't fix too many cells, a 1-3 mm pellet } in a 1.5 ml eppy is plenty. Of course your adding a 2x fix at equal } vol to the suspension to get the final concentration in step 1. Not } all samples will fix optimaly so you can play with fix conc. and time. } Microwave fixation is an option to cut time down. } Good Luck } Mike D. } }
I forgot to mention in my former query that the obvious method of weighing, drying, weighing again is not practical with these samples. They are too small and delicate to accurately removes excess water from the outside--the gross error is too large. Thanks Grace
POSTDOCTORAL POSITION IN ELECTRON MICROSCOPY AND MATERIALS SCIENCE OF CATALYTIC MATERIALS
Corporate Research Laboratory Exxon Research and Engineering Company.
The position will involve the use of high resolution and analytical electron microscopy for understanding model supported catalysts, and for developing new catalysts used in refining or chemical production. Requirements include expertise with high resolution electron microscopy, electron holography, field-emission analytical microscopy, and scattering physics. Experience with the materials science of catalysts is desirable for this rapidly advancing research program. Research in this area involves a multidisciplinary team approach which provides an excellent learning environment. Our laboratory offers advanced instrumentation for structure-property studies of catalysts, including a new field-emission TEM equipped with a rotatable electron biprism for electron holography. We have extensive software and computer systems available for the computational component of this research. Candidates should have experience with digital imaging using CCD systems, and experience with or knowledge of the analytical methods for processing electron holograms.
The term of the position is one year beginning early in 1996 with a likely extension to two years. Our research lab in rural New Jersey offers convenient access to Philadelphia, Princeton and New York City. Exxon offers an excellent working environment, salary commensurate with skills and experience, and excellent benefits.
Applicants for this position who meet the majority of the qualifications outlined above should forward a resume, publication list and two or three letters of reference to:
Dr. Mark M. Disko Corporate Research Laboratory Exxon Research and Engineering Company 1545 Route 22 East Annandale, New Jersey 08801
(908) 730-2503 FAX (908) 730-3314
Please do not respond directly to this list server. In the event you need further information rapidly, forward electronic mail to me at mmdisko-at-erenj.com .
} From: kennedy-at-nsi.edu (grace kennedy) } Subject: tissue water content } } A colleague needs to accurately determine the water content of very small } tissue samples (fish embryos at various stages of development). What would } be the best way to do this? Thanks for any suggestions. Grace }
Gross Analysis: Weigh, Freeze Dry Weigh again.
The difference in weights = H2O Content.
Regards,
Ed Monberg, GM em-at-mediacity.com LMDC (Laser Motion Development Company) 3101 Whipple Road Union City, CA 94587-1216 510-429-1060 voice 510-429-1065 fax
The recent mail threads concerning flatbed scanners and negative scanning provided very good information on the resolution and contrast requirements needed to satisfy various imaging formats. I would like to provide for informational purposes only technical specifications on two Nikon scanners that have not been mentioned.
If you do not wish this to view this information please don't download the attached file.
SCANTOUCH: $1,535 SRP =0D Gives Your Images That Professional Touch =0D If you spend a lot of time scanning, you'll appreciate Nikon's fast, versatile, and affordable high-resolution color scanner. Backed by Nikon'= s renowned design and quality, the ScanTouch flatbed scanner offers the hig= h level of performance and value you demand. Take for instance its outstand= ing 1,200 dpi resolution with a super-fast scanning method. And professional scanning software for both Macintosh and IBM computers. Plus other rich, image-enhancing features ideal for your professional-quality layouts and publications using both images and text. =0D High resolution: 1,200 x 1,200 dpi Featuring 24-bit full color with an optical resolution of 565 x 1,200 dpi= , the Nikon ScanTouch offers an impressive 1,200 x 1,200 dpi hardware interpolated resolution. If you require finer image detail for line art a= nd graphics, its integrated powerful driver software provides a software interpolated resolution of 2,400 by 2,400 dpi or higher. =0D 10-bit internal processing per color When it comes to detailed color reproduction, the Nikon ScanTouch makes n= o compromises. A 10-bit A/D conversion for each color (R, G, B) with ColorS= ync compatibility ensures that your photos and artwork are accurately reprodu= ced with fine color gradations. =0D Three scanning modes Because requirements vary from job to job, the ScanTouch offers three versatile modes for speed and quality (A4-size, 300 dpi full color): Norm= al Mode (approx. 60 sec.); Fast Mode (30 sec.); and High-Quality Mode (appro= x. 120 sec.). =0D Compatibility with your personal computer The ScanTouch comes ready to use with popular scanning packages - Photosh= op plug-in for Mac and PC and TWAIN compliant drivers (for PC only). =0D Fastest-class color preview Just as important to productivity as great image quality is being able to=
preview your scanned images quickly. At 12 sec. per page (A4), ScanTouch provides one of the fastest color preview times in its class. =0D Wide scanning area A wide surface scanning area of 8.5 x 14 in. lets you use the ScanTouch f= or most standard paper sizes such as A4, US Letter, and Legal. =0D Multifunctional, easy-to-use driver software Supports four types of output: line art, halftone, grayscale and full col= or. Available with automatic exposure, manual gamma compensation, manual gamm= a simulation, as well as cropping. You can also customize resolution and im= age specifications and adjust sharpness processing to emphasize contours and blurring processing to prevent moire effect when scanning printed materia= ls. =0D SCSI-II compatibility For fast transfer rate of image data to your computer, this high-quality scanner supports the advanced SCSI-II standard. =0D High character resolution for OCR applications When used with standard commercial OCR (optical character recognition) software, ScanTouch provides quick and accurate character reading. =0D Bundled software Comes with Caere's OmniPage Direct OCR software and Light Source's Ofoto,= an imaging application featuring automated scanning functions, color calibra= tion and image editing capabilities. (Note: Nikon will temporarily be providin= g Photoshop LE instead of Ofoto for the Macintosh model Scantouch. Ofoto wi= ll be made available at a later date, free of charge. However, use of Ofoto = will require a modification to Scantouch. There will be a service fee for this=
modification. Please contact Nikon's Service Department -at- 516-547-4351 fo= r further information). =0D Specifications - Original Document: Reflective original (printed/photo), max. 8.5 x 14 i= n.; Transparency or negative original (35mm to 5 x 5.7 in. film, at up to 1,200 dpi), Max. size with full rotation: 4 x = 5 in. - Reading Method: Fixed original, moving optical unit flatbed scanning - Sensor: CCD linear image sensor - Light Source: Fluorescent lamp - Color Separation: 3-pass RGB - Effective Reading Area: Reflective Original 8.5 x 14 in. (216 x 356mm);=
Optical Resolution: 565 x 1200 dpi; Interpolated Resolution: 1,200 x 1,20= 0 dpi (hardware), 2,400 x 2,400 dpi (software) - A/D Conversion: 10 bits/color - Output Data: Line Art, Halftone: 1-bit/pixel; Grayscale: 8 bits/pixel; = Full Color: 24 bits/pixel - Reference Reading Time (A4 300 -dpi Full Color): Approx. 30 sec. (Fast Mode, without exposure compensation) - Interface: SCSI (Complies with SCSI-II) - Dimensions (W x H x D): 14.8 x 6.3 x 23.6 in. (376 x 159 x 599mm) - Weight: Approx. 26.5 lbs. (12kg) - Power Requirements: AC 100 - 240V, 50/60Hz, max. 50W - Environmental Conditions - Temperature: +50xF to +95xF (+10xC to +35xC)=
=0D
Optional Transparency Unit AT-45: $675 SRP To scan transparencies and negative film (35mm to 5 x 5.7 in. film, at up= to 1,200 dpi), Nikon offers a low-profile, lightweight transparency unit tha= t's remarkably easy to use. =0D
All names of companies and products are trademarks or registered trademar= ks of their respective holders. =0D Specifications and equipment are subject to change without notice or obligation on the part of the manufacturer. June, 1994. =0D LS-4500AF Multi-format Film Scanner SRP: $11,295 =0D New Technology for Enhanced Productivity =0D * Supports formats from 35mm to 4x5 in. * Scanning head with dual-optical system * Precision 12-bit A/D conversion maintains wide dynamic range =
* Autofocus ensures consistently sharp scans * 360x-rotation film carriers with masks for all popular frame sizes streamlines production =
* Easy-access front-loading design simplifies film handling =0D One scanner for multiple formats =0D The new Nikon multi-format scanner can handle popular film sizes from 35m= m up to 4x5 in. It features 12-bit A/D conversion, 3000 dpi resolution on 35m= m and 1000 dpi on 4x5, and it's small enough to fit on your desktop! The LS-4500AF has an automatic energy-saving cycle and can handle your most demanding high-end film scanning requirements. =
=0D Dual-optical CCD scanning heads support multiple formats =0D Now, a single device can fulfill your multi-format needs. Nikon solved t= he challenge of matching film size to scanning engine by combining two scann= ing systems within the same device. The result is a scanner with high resolu= tion and performance, giving you the quality you need for all of its formats. =0D High-performance signal processing and mechanical accuracy =0D Preserving the wide dynamic range found on commercial transparencies, the=
LS-4500AF is ideal for professional applications. Color accuracy is tigh= tly controlled through 3D color matrix processing in hardware. The LS-4500's=
DSPs provide the power to scan and process any final size up to 3000 dpi = in 1 dpi increments, even from panorama formats! Its native scanner intellige= nce handles a wide array of positive and negative film types, formats, color masks and dye sets. =
The LS-4500's unique illumination path ensures a compact desktop footprin= t, and provides uniform edge-to-edge brightness, regardless of the film form= at, or its particular angle of rotation in the film carrier. High-definition=
optics overcome the problem of geometric distortion, enabling 3000/1000 d= pi high-quality output. With a precision-machined ballscrew drive, LS-4500A= F
delivers remarkably accurate color registration. =0D Easy-to-use front loading design =0D Film handling with the LS-4500AF is a streamlined operation cutting hours=
from your scanning production time. With a versatile carrier and mask se= t, film can be quickly mounted, rotated to a precise angle, and inserted int= o the autoloading slot. It is positioned by a special servo system to precisely locate the film in the optical path. You can be scanning withi= n seconds, confident that the alignment and focus are accurate. =0D High-speed scanning and preview =0D The LS-4500AF scans fast - 120 seconds for full-frame 35mm at 3000dpi, an= d 180 seconds for full-frame 4x5 at 1000 dpi. Prescan is only 30 seconds o= n 4x5 and 20 seconds on 35mm. In addition to precise servo-controlled manu= al focus, Nikon's autofocus takes care of precision focusing for you. This scanner is suited to large-volume pre-press scanning operations. LS-4500= AF can breeze through complex assignments, putting the profit back into your=
scanning operation.Easy-to-use Nikon software =0D The LS-4500AF is also an easy scanner to use, giving you the comprehensiv= e controls you need for professional results. Bundled with the scanner, Ni= kon introduces a completely re-architected Photoshopx plug-in for Mac OS, and=
TWAIN source for Windows users. The new Nikon Scan driver runs with any Photoshop or TWAIN-compatible image editing software, or standalone, usin= g the Nikon Control application shell. =0D Nikon Scan Driver Software Requirements =0D Operating System Mac OSr / Macintosh Windowsr Computer Platform 68xxx CPU without FPU, or PowerPCx CPU IBMr PC-AT compatibles (386, 486, or Pentiumx CPU RAM Plug-in requires a minimum of 2MB free RAM, Virtual Memory and Modern=
Memory Manager compatible. Image editing applications typically require = a minimum of 5-8MB RAM. With system memory requirements exceeding 2MB, tot= al recommended RAM should be greater than 16MB for productive scanning. TWAI= N module requires a minimum of 2MB free RAM, Virtual Memory compatible. Im= age editing applications typically require a minimum of 5-8MB RAM. With syst= em memory requirements exceeding 2MB, total recommended RAM should be greate= r than 16MB for productive scanning. Hard Disk Installation requires a minimum of 1MB free space. 300 MB or larger disk is recommended for scanning operation Installation requires a=
minimum of 1MB free space. 300 MB or larger disk is recommended for scan= ning operation Display 640 x 400 (or larger) full color (24-bit) display recommended 640= x 480 VGA (or larger) full color (24-bit) display recommended Interface SCSI-II ASPI compliant board supporting WINASPI.DLL OS Version System 7.0 or later, in English, German, French versions * MS-DOSr version 5.0 or later (requires enhanced mode) * IBMr-DOS version 5.0 or later (requires enhanced mode) * MSr-Windows version 3.1 or later (Win16 environment), English, German, French versions =0D Nikon Scan Driver Software Features =0D Scanner source selection, source image type selection, resizable dialog b= ox, resizable preview, autoexpose, autofocus, manual focus, crop, zoom, resolution, resize, fiducial reference scale, pixel address coordinate display, on screen densitometer, sharpening, analog exposure, analog colo= r balance, contrast, brightness, color balance, white point, gamma curve ed= it, histogram, black point, final scan, eject, instant screen update on densi= ty and color adjustment, interactive helpLS-4500AF Specifications =0D 1. Reading System/Optics =0D Film types: 4x5 in.; 40mm, 65mm, 75mm, and120/220 formats, including 6x4.= 5, 6x6, 6x7, 6x9, up to panorama formats; 35mm film (single frame, 6-frame strip, mounted film) Transparency, positive or negative, color or monochrome Reading resolution: 5000-pixel monochrome linear CCD x 2 [A]: 1000 dpi reading resolution [B]: 3000 dpi reading resolution Note: Dual-optical system modes are indicated by [A] and [B] in this docu= ment No. of pixels [A]: Maximum 5000 x 12000 pixels [B]: Maximum 5000 x 18000 pixels Effective scanning area [A]: 5 in. (Main scan) x 6 in. (Sub-scan) [B]: 42mm (Main scan) x 6 in. (Sub-scan) Light source: 12V- 20W halogen lamp Color separation: RGB frame sequential Film carrier: The following film formats are supported by three types of = film carriers, and masks are used to mount the film according to aperture/fram= e size. All carriers can be manually rotated for exact alignment during sc= an, except the 35mm 6-frame strip carrier; 4x5 in. sheet film; 6x4.5, 6x6, 6x= 7, 6x9cm cut frames; 35mm cut frames (three-up) 35mm mounted slides (four-up= ); 35mm x 6 frame strip Imaging optics: [A]: 8 lenses in 4 groups [B]: 6 lenses in 4 groups Autofocus: Contrast detection by CCD, focusing area selectable, manual focusing by software-controlled servo =0D 2. Scanning/Signal processing =0D Image scanning Three-pass RGB Main scan: 5000-pixel monochrome linear CCD [A]: 5000 pixels (hardware-interpolated to 10000) [B]: 5000 pixels Sub-scanning: stepper driven film stage [A]: 3 steps / line (2000 dpi) [B]: 3 steps / line (3000 dpi) Scan time: [A] Pre-scan: Approx. 30 seconds; Final scan: Standard mode - Approx. 210=
sec, Medium-speed mode - approx. 180 sec (-at- 1000 dpi, 4500 x 3600 pixels for approx. 46.3MB) [B Prescan: Approx. 20 seconds; Final scan: Standard mode - Approx. 200 s= ec, Medium-speed mode - approx. 120 sec (-at- 3000 dpi, 3900 x 2600 pixels for approx. 29MB) Scanning spatial density: Pixel density: [A]: maximum 2000 dpi interpolated from 1000 dpi [B]: maximum 3000 dpi Pixel size: [A]: 12.7um square [B]: 8.5um square A/D conversion: 12-bits per color channel Output data: 8-bits per color channel =0D 3. Data transfer =0D Panel indicators:READY, BUSY, and ERROR states indicated by LED Scanning software: Photoshop plug-in for Mac OS and TWAIN source for Wind= ows. NikonScan application supports Photoshop plug-in and TWAIN source on both= Mac OS and Windows platforms Interface: SCSI-II Image transfer: Three-pass RGB frame sequential Maximum transfer rate: 1 MB/sec, or better =0D 4. Operating conditions =0D Power requirements: 100 - 120VAC/200 - 240VAC; 0.8A/0.4A, 50/60Hz Environmental Temperature: 10xC - 35xC (50xF - 95xF) Relative Humidity: 30 - 85% (non-condensing) Dimensions and weight: =
295 (W) x 420 (D) x 250 (H)mm; approx. 13kg 11.6 (W) x 16.5 (D) x 9.8 (H) in.; approx. 28.7 lb. =0D Macintosh is a registered trademark and Mac OS is a trademark of Apple Computer, Inc. MS, MS-DOS and Windows are registered trademarks of Microsoft Corporation=
Photoshop is a trademark of Adobe Systems, Incorporated IBM is a registered trademark, and PowerPC is a trademark of Internationa= l Business Machines Corporation Pentium is a trademark of Intel Corporation =0D Contacting Nikon is Easy! =0D In the US, for product literature and the name and number of a dealer nea= r you, call 800-52-NIKON. For Marketing, Technical and Service support for=
Nikon Electronic Imaging Products in North, Central, and South America pl= ease contact Nikon EID personnel at: =0D Nikon Inc. Electronic Imaging Department 1300 Walt Whitman Road, Melville, NY 11747-3064 Attention: Marketing, Technical Support or Sales =
Phone: 516-547-4355 Fax: 516-547-0305 800: 800-52-Nikon America Online: Keyword Nikon CompuServe: Go Nikon =0D For all other regions, consult the list below to find the Nikon subsidiar= y closest to you: =0D Nikon Corporation Electronic Image Engineeering Division Fuji Bldg., 2-3 Marunouchi 3-chome, Chiyoda-ku, Tokyo 100, Japan Phone: 81-3-3216-1034 Fax: 81-3-3214-8193 =0D Nikon Canada Inc. 1366 Aerowood Drive, Mississauga, Ontario, Canada L4W 1C1 Phone: 416-625-9910 Fax: 416-625-0103 =0D Nikon AG Kaspar-Fenner Strasa 6, 8700 Kunacht/ZH, Switzerland Phone: 41-1-913-61-11 Fax: 41-1-913-63-63 =0D Nikon GmbH Tiefenbroicher Weg 25, 40472 Dusseldorf, Germany Phone: 49-211-9414-0 Fax: 49-211-9414-330 =0D Nikon UK Ltd. Nikon House 380 Richmond Road, Kingston-upon-Thames, Surrey, KT2 5PR, United Kingdom Phone: 44-181-541-4440 Fax: 44-181-541-4584 =0D Nikon France S.A. 191, rue du Marche Rollay, 94504 Champigny-sur-Mame, France Phone: 33-1-45-16-46-00 Fax: 33-1-45-16-00-33 =0D Anam Instruments Co., Ltd. 197-7 Guro 3-Dong, Guro-Ku, Seoul, Korea Phone: 82-2-460-5032 Fax: 82-2-861-4895 =0D Lin Trading Co., Ltd. 2F. No. 270, Sec 3., Nanking E. Road, =
Taipei, Taiwan, Republic of China Phone: 886-2-740-3366 Fax: 886-2-773-5577 =0D Shriro (H.K.) Ltd. 2nd Floor, Hutchison House, 10 Harcourt Road (G.P.O. Box 181), Hong Kong Phone: 852-2524-5031 Fax: 852-2810-6586 =0D Shriro (Singapore) Pte. Ltd. Shriro House 11 Chang Cham, Singapore 0315 Phone: 65-472-7777 Fax: 65-472-1792 =0D Shriro (Malaysia) SDN BHD Lots 22 & 24Julan 225, Section 51A, 46100 Petaling, Jaya, Selangor, Daryl Ehsan, Malaysia (P.O. Box 10571, 50718 Kula Lumpur) Phone: 60-3-774-9842 Fax: 60-3-775-2463/775-2400 =0D Maxwell Optical Industries Pty Ltd. Unit 27, Level A, 100 Harris Street Pyrmont N.S.W 2009 Australia Phone: 61-2-660-7088 Fax: 61-2-660-8739 =0D T.A. Macalister Limited Private Bag 92146, Aukland, New Zealand Phone: 64-9-303-4334 Fax: 64-9-309-6502
Best wishes for this new year, hope that you will receive nice pictures...
There is a guy here in Strasbourg, who wishes to have a look on his oligodentrocytes in culture by SEM. For these purpose he uses antibodies coupled to gold to label the cells.
My question is, what is the minimal size of the gold particles to have a chance to see them by SEM and the best way to prepare the sample (metallisation or not, carbon spputering, tension etc...) for SEM.
The SEM we have is a XL 20 from Philips.
Any related litterature is welcome as we are novice in this field.
TIA
Marc Schmutz
______________________________________
Dr. SCHMUTZ Marc Electron microscopy lab. IGBMC BP 163 F67404 ILLKIRCH Cedex France
There is a method of estimating water content of specimens if you can do TEM with X-ray microanalysis and work carefully; after the method of Ingram FD & Ingram MJ (1986) Cell volume regulation studies with the elctron microprobe. In: The Science of Biological Specimen Preparation for Microscopy and Microanalysis (eds. J-P Revel, T Barnard and GH Haggis) pp.139-146. SEM Inc. Chicago.
It means working on sections and may be too detailed for what you need.
I did something similar, using the chlorine peak of Spurr's resin to represent 100% hydration, and then lesser peak counts to represent replacement of the water by tissue components. Tissue chlorine is normally pretty labile and is lost in processing.
Keith Ryan PLymouth Marine laboratory Citadel Hill Plymouth PL1 2PB, UK
I am looking for advice and pointers in the realm of image transfer and manipulation, and I know this forum has a wealth of people's experiences available.
The department here has a number of instruments which can generate images of various sorts - optical (light microscope), element maps (SEMs/EMPA), secondary/back-scattered electron (SEMs/EMPA), isotope maps(SIMS).
Any one sample may pass through several of these instruments, so that the same area can be examined using different techniques. At the moment images are transferred as hardcopy photographs or plots. We are starting to investigate the possibilities of storing images electronically, transferring them between instruments by wire rather than as hardcopy, then redisplaying them and relocating areas of interest. (We are also going to have to consider the problem of longer term image archiving, which has recently been subject to useful discussion on the list.)
Obvious questions immediately occur:
In what format should the images be stored? TIFF seems to be a current common standard for this sort of work, but are there better alternatives? Are there any reviews/publications which would help? (We are also going to have to consider the problem of longer term image archiving, which has recently been subject to useful discussion on the list.)
Each instrument's specimen stage has its own particular coordinate system. Relating coordinate systems should be possible, given reference locations on the samples, but how to define such reference points - using distinctive features, or by marking the specimen somehow?
Other laboratories must be doing this sort of thing - how have they got on, what are the problems? If they could start again, how would they change things? What do we need to look out for, and take particular care of?
This posting is one of the first steps on what is clearly going to be a long term project. I look forward to the sort of interesting responses and discussion which commonly arise in this mailing list.
Norman Charnley
====================================================================== Dr. Norman Charnley Tel: +44 1865 272012 (Laboratory) Electron Microprobe Laboratory +44 1865 272053 (Office) Department of Earth Sciences Fax: +44 1865 272072 University of Oxford Oxford OX1 3PR, UK. E-mail:Norman.Charnley-at-earth.ox.ac.uk
We have a technical paper available on colloidal gold immunolabelling and SEM at our Web site. The URL is: http://www.mwrn.com/ebs/ebs.htm I hope you find it helpful. Steven Slap, Vice-President
Good morning. For your information, there is a society which I belong to that is called The Microscope Historical Society, 14 Tall Acres Dr., Pittsford, NY 14534 that can help you. Also, there are dealers which focus on antique microscopes . Contact Conrad Schure at 908.431.5191 about their next instrument fair in February at Hyatt Regency Hotel in New Brunswick, NJ.
--------------------------------------
Bob
Robert R. Wise, PhD Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
------------------ RFC822 Header Follows ------------------ Received: by qmgate_backup.anl.gov with SMTP;4 Jan 1996 13:52:54 -0600 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id MAA06365 for dist-Microscopy; Thu, 4 Jan 1996 12:57:17 -0600 Received: from VAXA.CIS.UWOSH.EDU (vaxa.cis.uwosh.edu [141.233.128.1]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id MAA06362 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 Jan 1996 12:57:15 -0600 by VAXA.CIS.UWOSH.EDU (PMDF V5.0-3 #11864) id {01HZM347X4NK001Y3K-at-VAXA.CIS.UWOSH.EDU} for Microscopy-at-MSA.Microscopy.Com; Thu, 04 Jan 1996 12:56:31 -0600 (CST)
Thank you for all the responses to my request regarding possible home for used light microscopes. Apparently, there is a market for reasonbly priced scopes. I received several names of people to contact and 20 requests (18 email, one phone and one fax) for the 12 scopes we have. Many of the respondents wanted more than one scope. I have kept all requests and will talk to our purchasing agent to see what can be done. I'll get back to you all when I know more.
Bob
Robert R. Wise, PhD Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
Hello, It seems that one of our EDS detectors consumption of LN2 has increased as of late. I am making the assumption that possibly the vacuum needs regeneration in the dewar. Does anyone know where one might acquire the fitting to interface a vacuum system to the dewar of a Kevex detector? Thank you in advance, Randy Nessler
} } } Obvious questions immediately occur: } } In what format should the images be stored? TIFF seems to be a current } common standard for this sort of work, but are there better alternatives? } Are there any reviews/publications which would help? (We are also going } to have to consider the problem of longer term image archiving, which has } recently been subject to useful discussion on the list.)
Well, for my two cents worth, my opinion is that format is almost irrelevant, except for a couple of points. The reason for this is that there are numerous utilities for moving between formats, both public domain (such as the pbmplus and netpbm libs) and commercial.
Some formats have standardized compression methods, and many don't, but even that doesn't really matter in most cases since you can compress things without relying on the format. For instance, you can use LZW compression as a part of the TIFF file, or you can save it unformatted and then compress the file using a compression routine. For my images, the difference in efficiency between the two approaches has been small. Certainly, some methods incorporate more efficient compression than others, but still, I have found the practical effect of, say, 45% compressions versus 40% compression to be of little importance for the number of images I manipulate. You can certainly obsess over each little bit of efficiency, and there are a number of religious arguments for a number of methods, but I have found the differences of little practical value for my archives.
With the differences between the formats of minimal interest to me (with a few exceptions listed below), then the most important thing to me becomes what *manipulation* software I use for which set of images. For instance, I have one package that can read and write TARGA files easily, but reads and writes TIFF images slowly and with occasional errors. For images I use this software on, I use TARGA files. A different program does a better job with Sun raster format.
I tend to save all my images in TARGA, but that is because it seems to be read and written faster and with fewer errors for the software I use. But, frankly, it really doesn't matter. I suggest you first decide on what software you plan to use to manipulate your images and *then* worry about format.
With all of this, there are a couple of things that become important quickly:
1) Make sure you can save your images with enough depth. I generally use 24-, 32- and 48-bit images. Thus, formats which do not support at least 24 bit "truecolor" representation are useless for me. On the other hand, if you are going to be happy with 8-bit images, then you have greater flexibility. I don't think there is a 24-bit GIF format, for instance, and relatively few formats will easily allow you go move to 32-bits (Targa supports it well).
2) Make sure you use lossless compression if you plan to do any image enhancement. Lossy compression, such as jpeg, can destroy an image for enhancement -- you end up enhancing the compression artifacts instead of the data. Jpeg and fractal compression are notorious for this.
Given these two caveats, then, I suggest you first look at the software you plan to use for image manipulation, and then choose a format that fits that software nicely.
we have a Phillips EM300 from 1971 to sell. It was well serviced throughout its existence in our department. It is perfectly suited for routine checking for biologist or medicine personal. There is a high resolution bridge and a standart goniometer available. We will be able to offer this microsope on a negotiational base. Transfer of the equipment must be handled by the buyer. For futher details contact me through email or by fax. DEADLINE is 31.01.96
Andreas Loewe
Guten Tag Freunde der Elektronenmikroskopie,
in unserer Abteilung ist ein Phillips EM300 aus dem Jahr 1971 zu verkaufen. Das Ger=E4t wurde w=E4hrend der gesamten Zeit durch einen Wartungsvertrag gepflegt und ist in sehr gutem Zustand. Dieses Mikroskop ist ideal f=FCr Routinearbeiten von Biologen und Medizinern. Es ist sowohl mit einer Hochaufl=F6sungsplattform als auch mit dem normalen Goniometer ausgestattet. Transport und die Transportkosten =FCbernimmt der K=E4ufer. Preis ist Verhandlungssache. F=FCr weitere Informationen schreiben Sie mich bitte unte= r meiner email-Adresse oder per Fax an. EINSENDESCHLUSS f=FCr Interessenten ist der 31.01.96
Andreas Loewe
______________________________________________________________ Andreas Loewe Tel: +49-228-550-355 University of Bonn Fax: +49-228-678-413 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
I can only emphasize what Bill Oliver wrote: Start with the basic question and ask at what precision you want to set up your digital lab. The acquisition precision (contrast resolution) of most microscopes (except some operation modes for confocal, see Jim Pawley's recent remarks at "RE: Flatbed scanners: 12-bit versus 8-bit") is far more than 8-bit. Most hardware and new software is already available for 12-16 bit graytone and color (per channel). If you plan for the future, start with 16-bit uncompressed raw data handling. Every thing else may change monthly. You may start using what is available now for handling } 8-bit data and expand as new software and hardware becomes available. And don't spend much money for a "good" printer, a HP LaserJet is just fine when expanded with one of the available high resolution boards. I started to put many comparative image data from my own experience in designing a "Precision Digital Imaging Laboratory" in a WWW page at http://panda.uchc.edu/htklaus/index.html. May be this is of help to you. Best regards Klaus
Regarding Randy Nessler's question about a valve to interface with the vacuum jacket of an EDS detector.
I have an old valve that is manufactured by Cryolab, 159 Santa Fe Rd., San Luis Obispo, CA. This is an old valve and the company may be gone/moved. I have no connection with this company whatsoever.
I too have a detector with high LN2 consumption and am considering doing the job myself. I assume that one would need the appropraite valve, a high vacuum station, leak detector and bakeout blank/tape. Is anyone familiar with the procedure for regenerating the EDS cyrostat?
Thanks, Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
We are thinking about a new printer and are looking at the Tektronix 340+ Phaser. The quote that we received lists costs associated with printing as "20% fill of color is $0.13 per page". Does anyone know the actual costs per page? Is this the maximum if "1% of color is $0.01 per page"? How much does it vary with size/color of image? What does this really mean? Anyone tried this machine yet?
Any/all help will be greatly appreciated. TIA.
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
After my reply about file formats, I got a bunch of requests about this mysterious pbmplus/netpbm library routine package.
Here's the story:
Way back when, there was this truly great dude by the name of Jef Poskanzer, who decided to write a little library to convert between formats. He decided to create his own simple, easily editable and readable ascii format for greyscale, binary, and color images. The idea was to concentrate on making the format simple to poke around in rather than being space-efficient since any image would be in the simple format only "in transit". This would make it easier for folk to write specific routines to convert their pet formats to and from these simple forms rather than worrying about trying to figure out every possible combination.
These simple formats were called Portable GreyMap, Portable BitMap,and Portable PixMap images respectively, or pgm, pbm, and ppm images. He and his buddies then wrote a ton of routines to go to and from these simple formats. Thus, for instance, to convert from GIF to TIFF, one would go from GIF to ppm using the routine giftoppm, and then from ppm to TIFF by the routine ppmtotiff.
A number of simple manipulation routines were added (rotation, scaling, smoothing, etc.) as well as some other sophisticated and useful things. In addition, a number of routines attempted to make the distinction between pgm, ppm, and pbm transparent, and rolled them into the "Portable aNyMap" or pnm format. Thus, one could also go from GIF to TIFF by using giftopnm and pnmtotiff routines without having to know if the image was greyscale or palette-based color. The pnm format wasn't really a new format, but was used to denote those routines which could accept pgm, pbm, and ppm format images and figure out which "real" conversion routine to call.
This work of public service, called the "pbmplus" library became too much of a time sink, and Jef Pokanzer stopped putting out updates in 1991. It nonetheless represents a public service effort right up there with GNU and the Fish disks. All Hail these folk.
After this work, a number of other community-minded authors added newer formats and other routines to the library. Since these sometimes did not conform to Pokanzer's structural and stylistic conventions and verification requirements, they represented a superset over the pbmplus library rather than an extension of it. This was labeled the netpbm library. Thus, new stuff like SGI formats, are found in the netpbm libraries, but not the pbmplus libraries.
I am including the beginning of the README file from the last distribution I have installed on my system. I am sure that things have been added since.
There are some formats, particularly the vector-based things, that are not handled. If you are using Wavefront, 3DS, DXF, and other CAD-type formats, you might want to look elsewhere. One such place is the old Avalon site. The Avalon site was a repository of 3D format convertors, models, etc. for 3D and VR software. This site, originally maintained at a US Government military base, was discontinued and was picked up by Viewpoint, a company which makes 3D model datasets. It can be found at http://www.viewpoint.com. Viewpoint has gotten mixed reviews for its stewardship; some have attempted to set up alternate repositories and others have applauded Viewpoint's effort if not it's accomplishment. I neither endorse nor criticise Viewpoint or the Viewpoint site, but simply state that it is still currently the most complete site for finding public domain 3D format conversion routines. If you are interested in the alternate sites, I suggest you look at the 3D news groups and mailing lists. It is a common topic of conversation on the Autodesk 3D studio newsgroup and list.
Sites for these and other routines can also be found in the FAQs for the graphics newsgroups.
One final note: Some of these routines are in turn dependent on other library distributions. For isntance, the tiff routines are dependent on the libtiff libraries which are a separate distribution. A libtiff library *is* included in the standard netpbm distribution, but the libtiff development history is rather independent. Thus, you will want to keep these libraries current if you want to be able to read and write newer flavors of tiff.
Now, here's the start of the netpbm README, which gives the sites as of early 1994 for the packages:
This README is dated 1993, but was current as of MARCH 1994.
N E T P B M Release 7 December 1993
Netpbm is a toolkit for conversion of images between a variety of different formats, as well as to allow a few basic image operations. The package is intended to be portable to many platforms. It has been tested under UNIX (BSD and SYSV, e.g. SGI, Sun4, Sun386i, DEC and Apollo DN 3500), VMS and Amiga OS. There are also compiler directives in it for MS-DOS.
You'll find the latest release of Netpbm at the following sites: * wuarchive.wustl.edu (128.252.135.4), directory /graphics/graphics/packages/NetPBM * ikaros.fysik4.kth.se (130.237.35.2), directory /pub/netpbm. * ftp.informatik.uni-oldenburg.de (134.106.1.9). This site also carries binaries for the Amiga. * peipa.essex.ac.uk (155.245.115.161), directory ipa/src/manip * ftp.rahul.net (192.160.13.1), directory /pub/davidsen/source * ftp.cs.ubc.ca, directory /ftp/archive/netpbm
You'll also find a mirror site at the BBS: * sixhub.tmr.com, phone +1 518 3468033, in the "source" area.
Hope you find this useful.
billo
Any opinions are mine, and not necessarily those of Uncle Sam, or any of his kith, kin, minions, or functionaries.
that format is almost } irrelevant, except for a couple of points. The reason for this is } that there are numerous utilities for moving between formats, both } public domain (such as the pbmplus and netpbm libs) and commercial.
} 1) Make sure you can save your images with enough depth. I generally } use 24-, 32- and 48-bit images. Thus, formats which do not support } at least 24 bit "truecolor" representation are useless for me.
and relatively few formats will easily allow } you go move to 32-bits (Targa supports it well). } } 2) Make sure you use lossless compression
Thanks, that you came forward and stressed the same points I try to emphasis.
A quick question to you: I started to do my digtal DH imaging on a Silicon Graphics workstation and of cause there is Tara File Format at home. However, most of my PC and Mac based image acquisition is still in MSA-Standard TIFF. You mentioned free software for file conversion. Could you please provide a source for me?
I'm preparing a presentation on polarized light microscopes that are equipped for epi-fluorescence illumination, and need information from microscope manufacturers regarding instruments purchased thus equipped and retro-fitted with vertical fluorescence attachments.
I'm assuming that microscope manufacturers monitor this list. Please reply directly to me. Thanks.
James Martin Director of Analytical Services and Research Williamstown Art Conservation Center Williamstown, MA
We have used critical point drying for many years in our laboratory to dry biological samples for SEM. Recently I decided to purchase some Peldri II to test as an alternative to critical point drying, however when I contacted Ted Pella they informed me that they are no longer selling it. Instead they are selling two substitutes: tetramethylsilane and hexamethyldisilazane. Does anyone have any experience with these yet? How do they compare to Peldri and critical point drying? Why was Peldri discontinued?
Jim Romanow Electron Microscopy Facility Physiology and Neurobiogy Department The University Of Connecticut Storrs bsgphy3-at-uconnvm.uconn.edu
______________________________________________ Preliminary Announcement: THIRD WORKSHOP ON INDUSTRIAL APPLICATIONS OF SCANNED PROBE MICROSCOPY May 2-3 1996 NATIONAL INSTITUTE OF STANDARDS AND TECHNOLOGY Gaithersburg MD 20899 ______________________________________________
A third Workshop on Industrial Applications of Scanned Probe Microscopy (IASPM) will be held at NIST on May 2-3 1996. The purpose is to continue exploring SPM standardization and development needs anticipated for the next decade, with a focus on industrial SPM users and hurdles that limit broader use of SPM for industrial problem solving.
For more information, contact: Dr. John Dagata National Insitute of Standards and Technology 220-A117 Gaithersburg MD 20899 301-975-3597 voice 301-869-0822 fax john.dagata-at-nist.gov
Are there any EM people heavy into Image processing and analysis. Currently I am using a Kontron IBAS (version 2.0) and a KS400 Image analysis system. Soon we will be incorporating a Telepathology system for communications between the US, Switzerland, and Japan. Current Image imput for our routine analysis come from EM micrographs, stored images, Kodak digital camera, from the light microscope, and any source that allows us to capture, store, and process images. We have several video camera, including a Sony 960 color camera, B&W camera. We currently use PAL and NTSC formats.
I would be interested in knowing how people use computerized Image Analysis for Stereology and Morphometry. Would be interested in knowing how to get around point counting techniques with Electron Micrographs.
I would be willing to share IBAS macros with interested parties. Gregory Argentieri Sandoz Electron Microscopy lab East Hanover, NJ 07936 201-503-8617 Greg2NJ-at-AOL.COM
Would welcome interested parties to exchange macros, ideas, image processing tips etc.
Are there any EM people heavy into Image processing and analysis. Currently I am using a Kontron IBAS (version 2.0) and a KS400 Image analysis system. Soon we will be incorporating a Telepathology system for communications between the US, Switzerland, and Japan. Current Image imput for our routine analysis come from EM micrographs, stored images, Kodak digital camera, from the light microscope, and any source that allows us to capture, store, and process images. We have several video camera, including a Sony 960 color camera, B&W camera. We currently use PAL and NTSC formats.
I would be interested in knowing how people use computerized Image Analysis for Stereology and Morphometry. Would be interested in knowing how to get around point counting techniques with Electron Micrographs.
I would be willing to share IBAS macros with interested parties. Gregory Argentieri Sandoz Electron Microscopy lab East Hanover, NJ 07936 201-503-8617 Greg2NJ-at-AOL.COM
Would welcome interested parties to exchange macros, ideas, image processing tips etc.
We have a Hitachi HU-11E (~1968) which has to make way for new developments. It was in regular use until about 2 years ago, now needs reference batteries and a kick-start. If anyone has a use for spares or the complete unit then please make contact with us ASAP. No reasonable offer refused !! It has to be cleared by March.
Tony Bruton Electron Microscope Unit University of Natal Pietermaritzburg Kwa-Zulu Natal South Africa Phone 0331 2605155 or fax 0331 2605776
If you are in need of vaccum pumping of an EDS detector, you might give contact;
Doug Conners TN Analyzer Service 608-798-2005
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
Message-Id: {199601101604.KAA28262-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Bob -
There is, or was, a wonderful shop called Historical Technology in Marblehead, Mass. that buys and sells old microscopes as well as other pieces of technology. I haven't delt with them in a long time, and they may no longer be in business. But it would be worth the effort to contact them just to get one of their catalogs. They are:
Historical Technology 6 Mugford St. Marblehead, Massachusetts 01945 (617) 631-2275
Sorry that this reply is so late, but I had to go mining in a closet to find the catalog.
Joiner ************************************
At 02:07 PM 1/4/96 +0000, you wrote: } Our department has about a dozen old Spencer light microscopes--monoccular, } black laquer with lots of brass, probably pre-WWII. Does anyone know if } there is a market for such things out there? Are there 'antique' } microscope dealers who might be interested in these? They are not very } functional but too nice to throw away. } } Bob } } } Robert R. Wise, PhD } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-vaxa.cis.uwosh.edu } } } }
Jim Romanow asked: } We have used critical point drying for many years in our laboratory to dry } biological samples for SEM. Recently I decided to purchase some Peldri II } to test as an alternative to critical point drying, however when I } contacted Ted Pella they informed me that they are no longer selling it. } Instead they are selling two substitutes: tetramethylsilane and } hexamethyldisilazane. Does anyone have any experience with these yet? How } do they compare to Peldri and critical point drying? Why was Peldri } discontinued? }
Peldri II was dropped because it is a Freon - restrictions are becoming tighter - and because Pelco is an environmentally conscious company (I have no financial or emotional connection to Pelco). In our facility, we have used HMDS and it gives similar results as Peldri and CPD. Since your results will be highly dependent upon the type of specimen and processing (fixation, dehydration, etc) it is best to run a comparative trial using HMDS, TMS and CPD. I suspect that HMDS will be fine. Contact me if you need more info.
############################################################################# John J. Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
I posted this some time ago and got not a single response. Someone here approached me about a fixative called K11 that I never used. Please let me know what you know about it or give a reference for starter. Thanks. ****************************************************************** *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * * {Fermin-at-tmc.tulane.edu} Prof. Pathology & Otolaryngology * * http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html * ******************************************************************
} We have used critical point drying for many years in our laboratory to dry } biological samples for SEM. Recently I decided to purchase some Peldri II } to test as an alternative to critical point drying, however when I } contacted Ted Pella they informed me that they are no longer selling it. } Instead they are selling two substitutes: tetramethylsilane and } hexamethyldisilazane. Does anyone have any experience with these yet? How } do they compare to Peldri and critical point drying? Why was Peldri } discontinued? } } Jim Romanow } Electron Microscopy Facility } Physiology and Neurobiogy Department } The University Of Connecticut } Storrs } bsgphy3-at-uconnvm.uconn.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
We frequently use HMDS instead of critical point(CPD) It does not work well on all samples. Plants generally require CPD. Many microbes, insects, cells cultures and animal tissues can be successfully dried via HMDS. We only briefly tested Peldri and were not impressed. It showed no advantage over HMDS and was more tedious to use. We have not tried the Tetramethylsilane, but I suspect it would perform much the same as HMDS. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
} We have used critical point drying for many years in our laboratory to dry } biological samples for SEM. Recently I decided to purchase some Peldri II } to test as an alternative to critical point drying, however when I } contacted Ted Pella they informed me that they are no longer selling it. } Instead they are selling two substitutes: tetramethylsilane and } hexamethyldisilazane. Does anyone have any experience with these yet? How } do they compare to Peldri and critical point drying? Why was Peldri } discontinued? } } Jim Romanow } Electron Microscopy Facility } Physiology and Neurobiogy Department } The University Of Connecticut } Storrs } bsgphy3-at-uconnvm.uconn.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
We frequently use HMDS instead of critical point(CPD) It does not work well on all samples. Plants generally require CPD. Many microbes, insects, cells cultures and animal tissues can be successfully dried via HMDS. We only briefly tested Peldri and were not impressed. It showed no advantage over HMDS and was more tedious to use. We have not tried the Tetramethylsilane, but I suspect it would perform much the same as HMDS. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
} A colleague needs to accurately determine the water content of very small } tissue samples (fish embryos at various stages of development). What would } be the best way to do this? Thanks for any suggestions. Grace
Dear Grace, Can your colleague measure the weights and/or volumes of both the hydrated and lyophylized embryos? ESEM could give you the hydrated volumes, and coating the residue and using SEM might be adequate for the dehydrated volume as long as the residue packs with no voids. It might be easier simply to weigh both specimens. I realize, of course, that volatile salts will be lost during lyophylization, so that may not give accurate enough results. On another tack, is there a measure--such as osmotic activity-- which could be used? If so, one could put the embryos in a solution of PEG or some other macromolecular or impermient solute and note the concentration at which the embryos neither swell nor shrink. That might give enough infor- mation to determine the water content if there are a series of tissue samples of differing contents to use as standards. Good luck. Yours, Bill Tivol
Message-ID: {199601101405.JAA22696-at-IndyNet.indy.net} To: SPM List {spm-at-di.com} , Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
Dear Lynn:
I got your round robin e-mail about Scanning 96. I am submitting two papers but find there is an ambiguity in the instructions for preparing the abstracts. Please advise ASAP tp pe13-at-cus.cam.ac.uk whether the abstract should be only about 750 words long OR whether I can write an abstract which nearly fills the two pages on the Instructions to Authors. I would prefer the latter as I have a lot I want to cram in. Rest assured the two abstracts will be wih you by the end of the month OR when you have dug your self out of the snow drifts
Best wishes
Patrick Echlin
On Wed, 10 Jan 1996, Lynn Savino wrote:
} To all those interested in participating in SCANNING 96 } } Deadline for abstracts is February 1, 1996. }
} We have used critical point drying for many years in our laboratory to dry } biological samples for SEM. Recently I decided to purchase some Peldri II } to test as an alternative to critical point drying, however when I } contacted Ted Pella they informed me that they are no longer selling it. } Instead they are selling two substitutes: tetramethylsilane and } hexamethyldisilazane. Does anyone have any experience with these yet? How } do they compare to Peldri and critical point drying? Why was Peldri } discontinued? } } Jim Romanow } Electron Microscopy Facility } Physiology and Neurobiogy Department } The University Of Connecticut } Storrs } bsgphy3-at-uconnvm.uconn.edu } } } Jim, We have used HMDS(hexamethyldisilazane) in the past with good result. I still like to CPD the non-repeateble samples but have found HMDS to be suitable for most samples. Hope this helps
} It seems that one of our EDS detectors consumption of LN2 has } increased as of late. I am making the assumption that possibly the vacuum } needs regeneration in the dewar. Does anyone know where one might acquire } the fitting to interface a vacuum system to the dewar of a Kevex detector? } Thank you in advance,
Dear Randy, When this occurred with our Kevex, after a few times sending the whole thing back for regeneration, I had our shop make a back plate with a 1/2" copper tube attached. A swagelok fitting and valve allowed me to attach to the column vacuum, and I've been able to use that for the rege- neration ever since. The resolution is still reasonable after } 10 years-- 155 ev, as I remember--so this method works for us. If you have a very clean column vacuum, you can attach a turbo or ion pump (possibly a cold- trapped DP will do, but ask Kevex) to the tubing and not foul up the column. ~10^-6 or so will regenerate the cryosorbant. Good luck. Yours, Bill Tivol
hello fellow digital microscopists. from snow bound Philly comes a query for advice on color image processing of images. does anyone use a package that can quantify/image process color data instead of just greyscale?
please post replies directy and I will share summary with all.
._____________________ | Peter J. Hahn | ------------- | Thomas Jefferson University 3401 Spring Garden Street Philadelphia, PA. 19104 tel : 215-349-0099 | fax : 610-356-3451 | peter.hahn-at-mail.tju.edu | -------------+-----------------+----+------+
The Marine Biological Laboratory in Woods Hole, MA has announced the following course. More information and applications can be obtained at http://www.mbl.edu/html/guide/guide.home.html or by writing admissions-at-mbl.edu
Cell and Molecular Imaging Using Cryotechniques
Courses Dates: May 28-June 5, 1996
Course Description: An eight-day comprehensive course/workshop on cryotechniques for cell and molecular imaging and analysis. Lecture and special discussion sessions in the mornings and hands-on lab sessions in the afternoons and evenings will form the heart of the course. Limited to 24 participants.
Topics will include freezing methods (including high pressure freezing), freeze-substitution, cryosectioning, immunogold localization, freeze-fracture, Low Temperature SEM, cryo microscopy (TEM), image processing and X-ray microanalysis. Lectures and labs will be conducted by experts in the field from universities and industry.
Attendees will be expected to have a basic knowledge of electron microscopy. The class will attend common lecture sessions and will divide into groups for lab sessions and discussions on special topics. Attendees will be encouraged to consult with the course director about bringing their own specimens for use during lab sessions.
Director: M.V. Parthasarathy, Cornell University.
Faculty: Frank Booy, National Institutes of Health; Patrick Echlin, Cambridge University, UK; Kent McDonald, University of California, Berkeley; Martin Mueller, ETH, Zurich, Switzerland; Klaus-Ruediger Peters, University of Connecticut Health Center; E. B. Prestridge, Princeton Gamma-Tech, Inc.; John Rash, Colorado State University; Daniel Studer, University of Bern, Switzerland; William Wergin, ARS-USDA; Karl Zierold, Max-Planck Institute for Molecular Physiology, Germany; and others to be named.
=-=-=-=-=-=-=-=-=-=-=-=-=-=-=- David Remsen Marine Biological Laboratory
You need to be a bit more specific. When some folk talk about "image processing," they mean the kind of things you do to get an image ready for presentation or publication -- annotation, a little contrast buffing, etc. Other folk mean playing with point spread functions and such for image regularization. Still others mean doing image manipulation for data extraction -- segmentation, object recognition, metrics of various sorts, etc. Finally, 2D and 3D products are rather different.
The type of package you might want depends upon the specific task and the platform you like. For instance, in my work with forensic image processing, I use AVS (Advanced Visualization Systems) as a programming environment, and incorporate libraries and code from a number of academic insititutions (for instance, DIAL and /usr/image from University of North Carolina and the University of Utrecht). AVS, and its big brother AVS Express, use a dataflow paradigm which takes the UNIX pipe to extremes. In this paradigm, you program little modules, say fft, and then plunk an icon for that module on the screen. You then connect other modules to it (say, a "read image" module) using little pipes. You can then build up larger applications by linking these little modules in networks. AVS comes with a set of supported modules, and there is a large set of community-donated public domain modules.
This is specific to UNIX-based systems, however. Most specific platforms have their own image processing libraries that you can buy. I know that Sun has something, SunVision, I think, and SGI also has its image processing toolbox. SGI also incorporates some image processing into its Explorer package, which uses the same kind of dataflow paradigm as AVS.
Excellent public domain Unix image processing packages which include advanced routines include Khoros from U. New Mexico, and IRAF. Khoros is both a toolbox and/or development environment which incorporates a dataflow paradigm much like AVS and Explorer. The AVS-like front end for Khoros is called Cantata. IRAF, which is the common platform for academic astronomical image processing, has some very nice tools for deconvolution and image restoration, but it is geared heavily towards astronomical models and is a true bear to install.
NIH has a package called, I think, NIH Image, but I have never used it. I have heard good things, but I think it is specific to Macs, is for greyscale only, and is 8-bit only.
For some of the cellular morphometric stuff, we tend to use PC-based software such as Optimas. There are, however, two or three other relatively good systems for doing metrics and more advanced image processing such as ImagePro and there's this French program by Noesis called, I think, Visilog. Each of these have routines for most common simple, intermediate and advanced image processing and 2D measurement tasks. I like the programming interface for Optimas. Visilog has, as far as I can see, a larger set of ready-to-use tools for mathematical morphologic manipulation, which you would expect from the French, I suppose. All of these systems allow, in one way or another, incorporation of home-brewed routines either as a link to compiled code or by using a macro language.
For annotation and aesthetic stuff, I prefer Picture Publisher by Micrografx. Many like Photoshop, and I have used it occasionally, but I still prefer Picture Publisher. Photostyler (Aldus) and Corel are also good products. I find Picture Publisher to be more intuitive and easier to use. It has a harder time successfully importing images, however. Our PCs are on a network with Unix boxes, and the images get to the PCs via nfs. My installation of Picture Publisher has problems often with reading large images over the network and with decoding large or compressed images. I often read an image from the network in using Photostyler, save it locally, and then manipulate it using Picture Publisher. I have found the Corel product a memory hog, and gets really slow with big (40 Mbyte) images.
Aside from the memory and importing hassles, the biggest differences with these artistic manipulators from that point on involve specific tools -- how you want to set brush sizes, how you want to play with overlays, how the gui is set up (whether you prefer type-in widgets, etc.). For instance, Picture Publisher has a slider transparency control on a toolbar for interactively playing with overlays, and allows you to play with overlay parameter during a paste. Other programs require multilevel menu selection to set transparency, which is not as easy. The (rather old) version of Photostyler I have used has a pretty primitive file requestor for loading images. Picture Publisher has a requestor that does thumbprints. This is a mixed blessing with a heterogeneous network -- it makes browsing easier, but the thumbprints can eat up disk space, they can "get lost" when the unix directories get mapped to DOS names, etc. Which set of widgets and approaches is most intuitive or easiest to learn is a matter of personal taste. Fractal Paint and associated tools have really nice interfaces for doing true "artistic" manipulation in the sense of having results that look like true artistic media (charcoal, oils, etc.) and "natural" textures. There is another package called {something - I forget} Matisse, which also attempts the "true" artistic approach. The {something} Matisse product, however, can get really slow with larger images. I have found it useless for work but a gas to play with.
There are a number of public domain systems which do limited image manipulation -- scaling, rotating, cropping, etc. Some of these have their own little areas of surprise. I have already discussed the pbmplus/netpbm libraries. There are others, such as ImageMagick, Utah Raster Toolkit, etc. which provide similar routines.
For integration of images into presentations and some buffing for publication (cropping, playing with backgrounds, etc.) I use Corel, though many also like Harvard Graphics. I just happened to have Corel at home, and never wanted to bother with the learning curve for Harvard Graphics or PowerPoint. My wife uses Harvard Graphics and I have helped her on occasion -- it is fairly intuitive.
For 3D modeling work, I have worked with AutoCAD, 3d Studio, and Lightwave 3D. Each has specific strengths and weaknesses, but I bet you aren't interested in these, so I will spare you reviews. For high-end production on UNIX boxes, there are Wavefront, Alias, SoftImage, and ElectroGig. I have worked with Wavefront and Gig, if you want reviews.
Finally, there are specific tools for such things as texture generation, 3D painting, titling, etc. For PCs, for instance, Kai's Power Tools is an excellent product. For UNIX boxes, Xaos has recently released a number of good tools.
3D visualization tools for medical images which incorporate some measurement capability include some commercial products which have been discussed ad-nauseum on the confocal list, so I won't discuss them here. There is a pseudo-commercial product out of Mayo called Analyze which is primarily a 3D visualization tool with some measurement capability. Last I heard, Rich Robb was charging about 5000 bucks for it. There is a relatively cheap tool from UPenn (Jay Udupa's lab) called 3DViewnix which runs about a thousand bucks which again is primarily a visualization tool with some measurement capability. There is a free tool out of Arie Kaufmann's lab at SUNY called VolVis which is, again, primarily a visualization tool with some measurement capability. I have heard good things about a program called Confocal Assistant, but have never seen or tried it. I believe it is freely distributable, but I am not sure.
hope this helps,
billo
All opinions are my own and do not necessarily reflect those of the US Govt, Army, Uncle Sam, yada, yada, yada.
On Thu, 11 Jan 1996 HAHNP-at-JEFLIN.TJU.EDU wrote:
} hello fellow digital microscopists. } from snow bound Philly comes a query for advice on color image processing of } images. does anyone use a package that can quantify/image process color data } instead of just greyscale? } } } please post replies directy and I will share summary with all. } } } } ._____________________ } | Peter J. Hahn } | ------------- } | Thomas Jefferson University } 3401 Spring Garden Street } Philadelphia, PA. 19104 } tel : 215-349-0099 | } fax : 610-356-3451 | } peter.hahn-at-mail.tju.edu | } -------------+-----------------+----+------+ } }
Our facility has been requested to examine a series of artificial heart valves made of high-tech graphite utilizing the SEM. We will need to initially examine the valves prior to the first test to establish a baseline of the surface. They will be returned to the testing site for further testing for wear then returned to us for re-examination. We will be unable to sputter coat the valves with any type of conducting material.
I would like any suggestions for proper examination of graphite. Since this is my first experinece with graphite, my first choice would be to examine the valves at a low Kv (5, 10, or 15) and at a long working distance of 39. My instrument is a JEOL JSM-35 SEM.
Any suggestions and/or comments may be forwarded to my direct e-mail address: aandrews-at-fdant.nctr.fda.gov Thank you.
Thanks to all who responded. _______________________________________________________________________________
Cesar, I use an aldehyde-based fixative called 'KII'. It is a dilute Karnovsky's fixative containing: 2% formaldehyde, 2.5% glutaraldehyde, 0.025% calcium chloride in 0.1 M sodium cacodylate buffer, pH 7.4.
Gary Login
In message {n1390845623.57279-at-msmail.tmc.tulane.edu} "Fermin, Cesar" writes: } I posted this some time ago and got not a single response. Someone here } approached me about a fixative called K11 that I never used. Please let me } know what you know about it or give a reference for starter. Thanks. } ****************************************************************** } *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * } *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * } *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * } *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * } * {Fermin-at-tmc.tulane.edu} Prof. Pathology & Otolaryngology * } * http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html * } ****************************************************************** }
Gary R. Login, D.M.D., D.M.Sc. Assistant Professor of Oral Pathology Beth Israel Hospital Department of Pathology 330 Brookline Avenue Boston, Massachusetts, 02215
Dear Colleagues, I recall recently people "advertizing" used optical microscopes on this list. I didn't need one at the time, so I did not save the messages, but I now find myself in need of a optical microscope. I specifically need one that has a calibrated focus knob so that I can make thickness meaurements in the 1-50 micron range by focusing on the front and back surfaces of our (transparent) samples. Please contact me if you know where I might find an inexpensive (used is fine, especially if it's inexpensive) microscope for the above task.
Has anyone run into a problem printing grayscale images on a Kodak/Codonics printer?
Ours have started looking tinged with green/yellow right out of the printer. I have read that the formulation of the ribbons may have changed and that the result is this sick green color.
I would like to avoid swapping color/grayscale ribbons if possible but my users don't like the color cast to their pictures. Any ideas for a solution?
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 jmkrupp-at-cats.ucsc.edu
I was leafing through Macworld the other day and realized that they had reviewed several scanners in the last 6 months. The recommendes ones were (4 or more stars out of 5):
Flatbed scanners:
HP Scanjet 3c 30 bit dynamic range, 600 dpi optical resolution, list $1179, street ~$700 208-323-2551
PixelCraft Pro Imager 8000 30 bit, 1200 dpi optical, list $13k 510-562-2480
35 mm negative scanner:
Polaroid SprintScan 35 30 bit, interpolated 2700 dpi, list $2495, street ~$1600
-Kirk _____________________________________________ Kirk Rogers krogers-at-materials.ecn.purdue.edu OR kirk.a.rogers.1-at-purdue.edu Purdue University, School of Materials Science and Engineering, 1289 MSEE building, W. Lafayette, IN 47907-1289 OFFICE: 317-494-8751 FAX: 317-494-1204 http://materials.ecn.purdue.edu/~krogers
} NIH has a package called, I think, NIH Image, but I have never used it. I } have heard good things, but I think it is specific to Macs, is for } greyscale only, and is 8-bit only.
I believe NIH Image supports 4, 8 and 16 bit TIFF (as well as PICT, pics, macpaint) images in both color and grayscale and 24 bit color images. Image will also open, edit, compare frames, average frames, etc. on captured video or quicktime movies. NIH Image now (v. 1.59) includes several built-in fast fourier transforms, although these run rather slowly on 68k Macintoshes. And heck, it's free!!
{ftp://zippy.nimh.nih.gov/pub/nih-image/}
The 8-bit part is in the way Image handles adjacent pixels. In other words, all contiguous pixels are counted during operations, not just the 4 pixels (4-bit) that share a side with the pixel in question. In doing microstructural analysis of some acicular structures we found our $50k workstation-based image analysis system was not able to analyze the microstructure as well as NIH Image because it was a 4-bit system.
For those interested in doing 2D image analysis and reconstruction, The National Center for Supercomputing Applications (NCSA) at UIUC, has a program called NCSA Image. I just visited their web site,
{http://www.ncsa.uiuc.edu/General/NCSAHome.html}
and found that Image is now called Collage, and is available for Macintosh and X-Windows for FREE. Collage uses a proprietary data format, HDF, but you can get Import2HDF, which will convert FITS, TIFF, GIFF, Pict, ascii and rastor files to HDF. NCSA Collage also supports real time interaction between scientists working on the same data sets at remote locations according to the readme file.
NCSA GelReader, which automates the measurement of DNA length using digitized electrophoretic gels is also available for free from NCSA, and is only available for the Macintosh.
(Insert standard disclaimer about not being intimately involved with these products here.)
-Kirk _____________________________________________ Kirk Rogers krogers-at-materials.ecn.purdue.edu OR kirk.a.rogers.1-at-purdue.edu Purdue University, School of Materials Science and Engineering, 1289 MSEE building, W. Lafayette, IN 47907-1289 OFFICE: 317-494-8751 FAX: 317-494-1204 http://materials.ecn.purdue.edu/~krogers
In response to recent discussion on this listserver regarding image analysis, I am forwarding description of newly available, FREE image analysis software, that was specifically designed to do for Windows users what NIH Image does for Mac users, and much more. I recently saw a demonstration of it and was impressed. Most impressive is that it is available absolutely FREE. It can be downloaded on the web from the Dental Diagnostic Science Dept of the University of Texas Health Science Center at San Antonio (ddxds.uthscsa.edu). This program was released the same day as Windows95 & has already been distributed worldwide. (This is not a commercial message. Even the creators of the software are not benefitting financially from its distribution). Soon to be released is a portion of the program that will make possible quantitation of the gels that molecular biologists love to run. =20 } } Date: Thu, 11 Jan 1996 16:03:08 -0500 (CDT) } From: "S. Brent Dove" {dove-at-uthscsa.edu} } Subject: Re: Image Analysis } To: "Nancy K.R. Smith" {smithn-at-uthscsa.edu} } } Hello Nancy, } } Here is a basic description of UTHSCSA ImageTool. Feel free to distribute this to any colleagues in the microscopy world. } } UTHSCSA } IMAGETOOL } } OVERVIEW } } UTHSCSA ImageTool (IT) is a free image processing and analysis program for Microsoft Windows 95=81 or Windows NT=81. IT can acquire, display, edit, analyze, process, compress, save and print gray scale and color images. IT can read and write over 22 common file formats including BMP, PCX, TIF, GIF and JPEG. Image analysis functions include dimensional (distance, angle, perimeter, area) and gray scale measurements (point, line and area histogram with statistics). ImageTool supports standard image processing functions such as contrast manipulation, sharpening, smoothing, edge detection, median filtering and spatial convolutions with user-defined convolution masks. IT also has built-in macro capabilities that allow the user to record repetitive tasks and playback saved macros to automate image analysis. } } ImageTool was designed with an open architecture that provides extensiblity via a variety of plug-ins. Support for image acquisition using either Adobe Photoshop plug-ins or Twain scanners is built-in. Custom analysis and processing plug-ins can be developed using the software development kit (SDK) provided (with source code). This approach makes it possible to solve almost any data acquisition or analysis problem with IT. } } ImageTool provides for geometric transformations such as rotate, flip vertical, flip horizontal and magnification up to four levels. All analysis and processing functions are available at any magnification factor. The program is a multiple document interface (MDI) application supporting any number of windows (images) simultaneously. =20 } } Spatial calibration is available to indicate real world dimensional measurements such as millimeters, microns, feet, miles, etc. for linear and area. Density or gray scale calibration can be done relative to radiation or optical density (OD) standards. } } IT version 1.1 now provides for object analysis and classification with over 20 morphological descriptors such as: area/perimeter, roundness, ferret diameter, compactness, major/minor axis length, centroid and many others. Any of these factors can be used automatically categorized and count objects within the image. } } ImageTool ver 1.1 supports the Data Translation DT3155 frame grabber for Windows NT. Other frame grabber board will be added in the coming months.= =20 } } ImageTool was written using Borland's C++ version 4.5 and the source code for the executable is available free of charge. IT was developed in the Department of Dental Diagnostic Science at The University of Texas Health Science Center, San Antonio, Texas. The program was developed by C. Donald Wilcox, S. Brent Dove, W. Doss McDavid and David B. Greer. } } } } } S. Brent Dove Voice: (210) 567-3333 } Diagnostic Sciences Fax: (210) 567-3334 } University of Texas Email: dove-at-uthscsa.edu } Health Science Center Web: ddxds.uthscsa.edu } San Antonio, TX USA ftp: maxrad6.uthscsa.edu } }
HAHNP-at-JEFLIN.TJU.EDU Once again, my opinions only:
You need to be a bit more specific. When some folk talk about "image processing," they mean the kind of things you do to get an image ready for presentation or publication -- annotation, a little contrast buffing, etc. Other folk mean playing with point spread functions and such for image regularization. Still others mean doing image manipulation for data extraction -- segmentation, object recognition, metrics of various sorts, etc. Finally, 2D and 3D products are rather different.
The type of package you might want depends upon the specific task and the platform you like. For instance, in my work with forensic image processing, I use AVS (Advanced Visualization Systems) as a programming environment, and incorporate libraries and code from a number of academic insititutions (for instance, DIAL and /usr/image from University of North Carolina and the University of Utrecht). AVS, and its big brother AVS Express, use a dataflow paradigm which takes the UNIX pipe to extremes. In this paradigm, you program little modules, say fft, and then plunk an icon for that module on the screen. You then connect other modules to it (say, a "read image" module) using little pipes. You can then build up larger applications by linking these little modules in networks. AVS comes with a set of supported modules, and there is a large set of community-donated public domain modules.
This is specific to UNIX-based systems, however. Most specific platforms have their own image processing libraries that you can buy. I know that Sun has something, SunVision, I think, and SGI also has its image processing toolbox. SGI also incorporates some image processing into its Explorer package, which uses the same kind of dataflow paradigm as AVS.
Excellent public domain Unix image processing packages which include advanced routines include Khoros from U. New Mexico, and IRAF. Khoros is both a toolbox and/or development environment which incorporates a dataflow paradigm much like AVS and Explorer. The AVS-like front end for Khoros is called Cantata. IRAF, which is the common platform for academic astronomical image processing, has some very nice tools for deconvolution and image restoration, but it is geared heavily towards astronomical models and is a true bear to install.
NIH has a package called, I think, NIH Image, but I have never used it. I have heard good things, but I think it is specific to Macs, is for greyscale only, and is 8-bit only.
For some of the cellular morphometric stuff, we tend to use PC-based software such as Optimas. There are, however, two or three other relatively good systems for doing metrics and more advanced image processing such as ImagePro and there's this French program by Noesis called, I think, Visilog. Each of these have routines for most common simple, intermediate and advanced image processing and 2D measurement tasks. I like the programming interface for Optimas. Visilog has, as far as I can see, a larger set of ready-to-use tools for mathematical morphologic manipulation, which you would expect from the French, I suppose. All of these systems allow, in one way or another, incorporation of home-brewed routines either as a link to compiled code or by using a macro language.
For annotation and aesthetic stuff, I prefer Picture Publisher by Micrografx. Many like Photoshop, and I have used it occasionally, but I still prefer Picture Publisher. Photostyler (Aldus) and Corel are also good products. I find Picture Publisher to be more intuitive and easier to use. It has a harder time successfully importing images, however. Our PCs are on a network with Unix boxes, and the images get to the PCs via nfs. My installation of Picture Publisher has problems often with reading large images over the network and with decoding large or compressed images. I often read an image from the network in using Photostyler, save it locally, and then manipulate it using Picture Publisher. I have found the Corel product a memory hog, and gets really slow with big (40 Mbyte) images.
Aside from the memory and importing hassles, the biggest differences with these artistic manipulators from that point on involve specific tools -- how you want to set brush sizes, how you want to play with overlays, how the gui is set up (whether you prefer type-in widgets, etc.). For instance, Picture Publisher has a slider transparency control on a toolbar for interactively playing with overlays, and allows you to play with overlay parameter during a paste. Other programs require multilevel menu selection to set transparency, which is not as easy. The (rather old) version of Photostyler I have used has a pretty primitive file requestor for loading images. Picture Publisher has a requestor that does thumbprints. This is a mixed blessing with a heterogeneous network -- it makes browsing easier, but the thumbprints can eat up disk space, they can "get lost" when the unix directories get mapped to DOS names, etc. Which set of widgets and approaches is most intuitive or easiest to learn is a matter of personal taste. Fractal Paint and associated tools have really nice interfaces for doing true "artistic" manipulation in the sense of having results that look like true artistic media (charcoal, oils, etc.) and "natural" textures. There is another package called {something - I forget} Matisse, which also attempts the "true" artistic approach. The {something} Matisse product, however, can get really slow with larger images. I have found it useless for work but a gas to play with.
There are a number of public domain systems which do limited image manipulation -- scaling, rotating, cropping, etc. Some of these have their own little areas of surprise. I have already discussed the pbmplus/netpbm libraries. There are others, such as ImageMagick, Utah Raster Toolkit, etc. which provide similar routines.
For integration of images into presentations and some buffing for publication (cropping, playing with backgrounds, etc.) I use Corel, though many also like Harvard Graphics. I just happened to have Corel at home, and never wanted to bother with the learning curve for Harvard Graphics or PowerPoint. My wife uses Harvard Graphics and I have helped her on occasion -- it is fairly intuitive.
For 3D modeling work, I have worked with AutoCAD, 3d Studio, and Lightwave 3D. Each has specific strengths and weaknesses, but I bet you aren't interested in these, so I will spare you reviews. For high-end production on UNIX boxes, there are Wavefront, Alias, SoftImage, and ElectroGig. I have worked with Wavefront and Gig, if you want reviews.
Finally, there are specific tools for such things as texture generation, 3D painting, titling, etc. For PCs, for instance, Kai's Power Tools is an excellent product. For UNIX boxes, Xaos has recently released a number of good tools.
3D visualization tools for medical images which incorporate some measurement capability include some commercial products which have been discussed ad-nauseum on the confocal list, so I won't discuss them here. There is a pseudo-commercial product out of Mayo called Analyze which is primarily a 3D visualization tool with some measurement capability. Last I heard, Rich Robb was charging about 5000 bucks for it. There is a relatively cheap tool from UPenn (Jay Udupa's lab) called 3DViewnix which runs about a thousand bucks which again is primarily a visualization tool with some measurement capability. There is a free tool out of Arie Kaufmann's lab at SUNY called VolVis which is, again, primarily a visualization tool with some measurement capability. I have heard good things about a program called Confocal Assistant, but have never seen or tried it. I believe it is freely distributable, but I am not sure.
hope this helps,
billo
All opinions are my own and do not necessarily reflect those of the US Govt, Army, Uncle Sam, yada, yada, yada.
On Thu, 11 Jan 1996 HAHNP-at-JEFLIN.TJU.EDU wrote:
} hello fellow digital microscopists. } from snow bound Philly comes a query for advice on color image processing of } images. does anyone use a package that can quantify/image process color data } instead of just greyscale? } } } please post replies directy and I will share summary with all. } } } } ._____________________ } | Peter J. Hahn } | ------------- } | Thomas Jefferson University } 3401 Spring Garden Street } Philadelphia, PA. 19104 } tel : 215-349-0099 | } fax : 610-356-3451 | } peter.hahn-at-mail.tju.edu | } -------------+-----------------+----+------+ } }
Received: from ibmstone.pku.edu.cn by pccms.pku.edu.cn with SMTP id AA07824 (5.67b8/IDA-1.5 for zqliu); Fri, 12 Jan 1996 09:50:26 +0800 Received: from sparc5.microscopy.com by ibmstone.pku.edu.cn with SMTP id AA06453 (5.67b8/IDA-1.5 for {zqliu-at-pku.edu.cn} ); Fri, 12 Jan 1996 10:25:00 -0600 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id MAA14626 for dist-Microscopy; Thu, 11 Jan 1996 12:16:11 -0600 Received: from tmcpop.tmc.tulane.edu (tmcpop.tmc.tulane.edu [129.81.48.30]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id MAA14623 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 11 Jan 1996 12:16:10 -0600 Received: from postoffice.tmc.tulane.edu by tmcpop.tmc.tulane.edu (AIX 3.2/UCB 5.64/4.03) id AA27752; Thu, 11 Jan 1996 11:42:41 -0600 Message-Id: {n1390759410.38417-at-msmail.tmc.tulane.edu}
**** Very Important !!!! My Telphone and Fax Numbers have been changed recently !!!
Zhen Quan Liu, Director, Ph.D Tel (86) 10 275 1427 (Office) Physics Building Tel (86) 10 275 3727 (Home) Electron Microscope Laboratory Fax (86) 10 275 1615 (Office) Peking University Beijing 100871, China E-mail(Office): zqliu-at-pku.edu.cn Email (Home) wl-at-ibmstone.pku.edu.cn
What exactly you want to know? _____________________________________________________________________________ Francisco Javier Hernandez Blazquez * Av. Prof. Lineu Prestes, 1524 Departamento de Histologia e Embriologia * 05508-900 Sao Paulo Instituto de Ciencias Biomedicas * e-mail fjhblazq-at-spider.usp.br Universidade de Sao Paulo * fjhblasq-at-biomed.icb2.usp.br ______________________________________________________________________________
I am looking for references which contain detailed information about how the magnification changes with working distance, spot size and voltage. On our Cambridge Stereoscan II, I have noticed a "curious" behaviour, that is, for a given set of microscope conditions, if we plot the real magnification as a function of nominal magnification, we get maxima and minima which seem reproducible. The range of magnification investigated is 100x to 10000x, no sample tilt, secondary electrons signal, linear micrometer with 10 micron spacings.
Thank you in advance for your help.
Antoine Ghanem e-mail address : ghanem-solvay-at-e-mail.com
Extra X400 information begins: Originator Name: Antoine (AGM) GHANEM Org Units: AC : NOH : LC-AN001 Organisation: NOHX400DEC Domain: BE/RTT/SOLVAY Node.Userid: IBMX400.184470
Message Id: 73820121106991/85925 MHS Importance: Normal Sent by Name: Antoine (AGM) GHANEM Org Units: AC : NOH : LC-AN001 Organisation: NOHX400DEC Domain: BE/RTT/SOLVAY Node.Userid: IBMX400.184470 Free Fmt Name: Antoine GHANEM Phone Number: 3422 Subject: SEM : magnification Recipients Name: INTERNET INTERNET Domain: GB/IBMX400/IBMMAIL Node.Userid: IBMMAIL.INTERNET Free Fmt Name: INTERNET INTERNET
I am looking for a small format (3-4" x 4-5" images, i.e. Polaroid size) grey scale printer, that is NOT restricted to screen dumps or Video dumps, but will function like a 'normal' printer output device. It can be parallel port, serial port or SCSI.
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
Message-Id: {199601121214.HAA16084-at-cpcug.org} X-Sender: jlibert-at-cpcug.org X-Mailer: Windows Eudora Version 1.4.3 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" William R Oliver {oliver-at-ipas4.afip.mil}
At 10:40 PM 1/11/96 -0500, Kirk Rogers wrote: } } NIH has a package called, I think, NIH Image, but I have never used it. I } } have heard good things, but I think it is specific to Macs, is for } } greyscale only, and is 8-bit only. } } I believe NIH Image supports 4, 8 and 16 bit TIFF (as well as PICT, pics, } macpaint) images in both color and grayscale and 24 bit color images. } Image will also open, edit, compare frames, average frames, etc. on } captured video or quicktime movies. NIH Image now (v. 1.59) includes } several built-in fast fourier transforms, although these run rather slowly } on 68k Macintoshes. And heck, it's free!! } } {ftp://zippy.nimh.nih.gov/pub/nih-image/} ----------------------------------------------------- Response: ** Actually, I believe that NIH Image currently cannot process color images and cannot acquire color images except as single 8-bit channels. Possible that someone has written some color extensions, but I am not aware of it. Check before buying a new computer. ------------------------------------------------------------------------------ } The 8-bit part is in the way Image handles adjacent pixels. In other } words, all contiguous pixels are counted during operations, not just the 4 } pixels (4-bit) that share a side with the pixel in question. In doing } microstructural analysis of some acicular structures we found our $50k } workstation-based image analysis system was not able to analyze the } microstructure as well as NIH Image because it was a 4-bit system. ----------------------------------------------------------------} Response: **This is not correct. The 4, 8, 24 bit designation describes the number of bits the program uses to represent the intensity value of each pixel, and has nothing to do with how the pixel neighborhoods are processed. The number of bits used to represent a numerical value in the computer is important to how well the system can represent the image information and derived information. The number of bits determine the range of values possible and specifies the power of 2 topping the range. For example, with 4 bits one can only represent 2^4 or 16 different values. With 8 bits the system can represent 2^8 or 256 values. Generally, color images are represented as three channels, each using 8-bits to quantize each pixel. Your old workstation may have also had algorithm deficiencies in doing neighborhood operations on images -- probably to save computation cost, but the main problem may have been in allowing only 4 bits to represent the entire dynamic range of your data.
** A second, but related, problem would arise if the system limits the bits assigned to represent derived images. The best systems will use floating point, or at least long integer, to represent intermediate values before the derived image. If you wanted to average images or do some transformation, you could run into serious problems if the values could be stored using 4 or even 8 bits. Once truncated, the image information is lost. ------------------------------------------------------------------------------ } For those interested in doing 2D image analysis and reconstruction, The } National Center for Supercomputing Applications (NCSA) at UIUC, has a } program called NCSA Image. I just visited their web site, } } {http://www.ncsa.uiuc.edu/General/NCSAHome.html} } } and found that Image is now called Collage, and is available for Macintosh } and X-Windows for FREE. Collage uses a proprietary data format, HDF, but } you can get Import2HDF, which will convert FITS, TIFF, GIFF, Pict, ascii } and rastor files to HDF. NCSA Collage also supports real time interaction } between scientists working on the same data sets at remote locations } according to the readme file. } } NCSA GelReader, which automates the measurement of DNA length using } digitized electrophoretic gels is also available for free from NCSA, and is } only available for the Macintosh. } } (Insert standard disclaimer about not being intimately involved with these } products here.) } } } -Kirk } _____________________________________________ } Kirk Rogers krogers-at-materials.ecn.purdue.edu } OR } kirk.a.rogers.1-at-purdue.edu } Purdue University, School of Materials Science and Engineering, } 1289 MSEE building, W. Lafayette, IN 47907-1289 } OFFICE: 317-494-8751 FAX: 317-494-1204 } http://materials.ecn.purdue.edu/~krogers } } } }
Hi, We are looking to purchase a used scanning microscope such as an AMR100 on which we can install a cold stage. We must take delivery and pay for the microscope by the end of March 1996. If anyone can tell us who could supply us with such a microscope this quickly please reply to me directly. We are located in Ottawa Canada. Thank you. Gisele Larocque larocqueg-at-em.agr.ca
} I specifically need one } that has a calibrated focus knob so that I can make thickness meaurements in } the 1-50 micron range by focusing on the front and back surfaces of our } (transparent) samples.
Dear Joyce, There are also thickness gauges which can do the job, if the samples are flat and the thickness uniform (some can also measure samples of non- uniform thickness). These may or may not be more expensive than used micro- scopes. Yours, Bill Tivol
Greg -- I have been using a PC based image analysis package from Imaging Research Inc.(no relation)to do morphometry and automated gold particle counting on digitized EM negs for a couple of years. The use of digital image analysis (IA) vs. point counting is highly dependant on your images and what information you want to extract from them. If the structures of interest can be easily segmented from the rest of the image, then IA is the way to go. But if the measurement involves structures which need a human eye and brain (still the best image analysis system avaliable) to pick them out then point counting may be a much more efficient method. An example of the latter would be measuring lengths of plasma membrane sub-domains or the limiting membrane of non-descript organelles. While you can trace the membranes using the cursor of an IA system and get good data, its VERY tedious. You can do alot more sampling with point counting techniques. If the specimen was immuno-gold labeled, then IA would work well to count the gold particles along those membranes since the computer can recognize the particles quite well because they are very dense and have a very circular profile. But I would still count them manually if there weren't too many. There are IA systems out there that allow you to use point counting methods on digital images. Imaging Reserch has an add-on to their systems that does stereology, but I haven't had a chance to try it out yet. I suspect that the ability to segment out the structures of interest will still be crucial, at least for automated measurements. But as long as you can manually define "hits" of the sampling grid on the specimen these systems may be pretty useful. Sorry for being long-winded, I'm alone in a snow-bound lab.
Greg Martin Dept. Cell Biology and Anatomy Johns Hopkins School of Medicine gmartin-at-welchlink.welch.jhu.edu On Tue, 9 Jan 1996 Gargent-at-aol.com wrote:
} Are there any EM people heavy into Image processing and analysis. Currently } I am using a Kontron IBAS (version 2.0) and a KS400 Image analysis system. } Soon we will be incorporating a Telepathology system for communications } between the US, Switzerland, and Japan. Current Image imput for our routine } analysis come from EM micrographs, stored images, Kodak digital camera, from } the light microscope, and any source that allows us to capture, store, and } process images. We have several video camera, including a Sony 960 color } camera, B&W camera. We currently use PAL and NTSC formats. } } I would be interested in knowing how people use computerized Image Analysis } for Stereology and Morphometry. Would be interested in knowing how to get } around point counting techniques with Electron Micrographs. } } I would be willing to share IBAS macros with interested parties. } Gregory Argentieri } Sandoz Electron Microscopy lab } East Hanover, NJ 07936 } 201-503-8617 } Greg2NJ-at-AOL.COM } } } } Would welcome interested parties to exchange macros, ideas, image processing } tips etc. }
Greg -- I have been using a PC based image analysis package from Imaging Research Inc.(no relation)to do morphometry and automated gold particle counting on digitized EM negs for a couple of years. The use of digital image analysis (IA) vs. point counting is highly dependant on your images and what information you want to extract from them. If the structures of interest can be easily segmented from the rest of the image, then IA is the way to go. But if the measurement involves structures which need a human eye and brain (still the best image analysis system avaliable) to pick them out then point counting may be a much more efficient method. An example of the latter would be measuring lengths of plasma membrane sub-domains or the limiting membrane of non-descript organelles. While you can trace the membranes using the cursor of an IA system and get good data, its VERY tedious. You can do alot more sampling with point counting techniques. If the specimen was immuno-gold labeled, then IA would work well to count the gold particles along those membranes since the computer can recognize the particles quite well because they are very dense and have a very circular profile. But I would still count them manually if there weren't too many. There are IA systems out there that allow you to use point counting methods on digital images. Imaging Reserch has an add-on to their systems that does stereology, but I haven't had a chance to try it out yet. I suspect that the ability to segment out the structures of interest will still be crucial, at least for automated measurements. But as long as you can manually define "hits" of the sampling grid on the specimen these systems may be pretty useful. Sorry for being long-winded, I'm alone in a snow-bound lab.
Greg Martin Dept. Cell Biology and Anatomy Johns Hopkins School of Medicine gmartin-at-welchlink.welch.jhu.edu On Tue, 9 Jan 1996 Gargent-at-aol.com wrote:
} Are there any EM people heavy into Image processing and analysis. Currently } I am using a Kontron IBAS (version 2.0) and a KS400 Image analysis system. } Soon we will be incorporating a Telepathology system for communications } between the US, Switzerland, and Japan. Current Image imput for our routine } analysis come from EM micrographs, stored images, Kodak digital camera, from } the light microscope, and any source that allows us to capture, store, and } process images. We have several video camera, including a Sony 960 color } camera, B&W camera. We currently use PAL and NTSC formats. } } I would be interested in knowing how people use computerized Image Analysis } for Stereology and Morphometry. Would be interested in knowing how to get } around point counting techniques with Electron Micrographs. } } I would be willing to share IBAS macros with interested parties. } Gregory Argentieri } Sandoz Electron Microscopy lab } East Hanover, NJ 07936 } 201-503-8617 } Greg2NJ-at-AOL.COM } } } } Would welcome interested parties to exchange macros, ideas, image processing } tips etc. }
Gooday all! I am seeking some help with locating a source of 'low viscosity nitrocellulose (LVN)'. I have used it some years ago to embed large specimens (approx. 10cm x 12cm x 10cm) for histology. LVN was obtainable from BDH but apparently they stopped selling it in 1991. I wish to find an alternative source of LVN or a suitable alternative embedding material. Any help would be appreciated. Thanks in advance.
Brett Cockman Brett W. Cockman Technologist in Charge School of Dentistry University of Western Australia 179 Wellington st., Perth, W.A. 6000 Voice: (619-2205834) Fax: (619-2213829) e-mail; bcockman-at-uniwa.uwa.edu.au
Lonely electrons: I think it was John Armstrong of Caltech who once pointed out to me that the number of microscope beam electrons in your TEM specimen at any one time is so small that the odds of such electrons interfering with each other to form diffraction patterns is quite small. Under some illumination conditions there may be no more than 1 beam electron in the column at a time! Hence diffraction patterns in the TEM are basically formed by individual electrons interfering with themselves! As you know, such interference will occur only if we don't take steps to determine the path of individual electrons through the specimen! If we look too closely at these paths, the diffraction patterns would disappear (cf. a Sci. Amer. ariticle in the past year or so on quantum erasure).
Fat electrons: The transverse coherence widths of electrons which make possible electron phase contrast (HREM) lattice imaging and probably electron holography might also be seen as lateral broadening of individual electron wave-packets via the uncertainty principle, which results because we know too much about their transverse momentum! My intuition tells we that we're talking about lateral wave-function spreads of say 15 Angstroms in a LAB6 HREM to more than 100 Angstroms in field emission gun system. Are these numbers reasonable? By increasing the spread of electron angles in the incident beam, this transverse coherence width can presumably be decreased (e.g. you want it small for Z-contrast imaging, I think), or varied as in the variable coherence-width strategies of Murray Gibson at U. of I.
Long electrons: The tight tolerences on high voltage stability and the emitted spread in electron energies means that our uncertainty in the longitudinal momentum of TEM electrons is quite small, and hence again by the uncertainty principle that the wave-packet spread in the direction of motion for TEM electrons can be quite large. Distances of say 1000 Angstroms come to mind! The associated tight distribution of incident electron energies decreases chromatic and instability damping of fine details in CTEM and HREM images, so that for most applications you may want your electrons "as long as possible". An exception might be in variable-coherence strategies (mentioned above), where shorter electrons might provide sensitivity to shorter-range vertical correlations.
Fast electrons: a back of the envelope calculation for 300 keV electrons gives gamma = (300+511)/511 = 1.587, so that they travel at w = c Sqr[1-(1/gamma)^2] = 0.777 c or [lightyears per inertial year] of elapsed time. However, if we consider traveler (i.e. electron or proper) time for such a speeding electron, this would give that they travel u = gamma*w = 1.232 [lightyears per traveler year] of elapsed time! With this spatial 4-vector velocity well over c, we're dealing with relativity in action! I wonder how many g's of acceleration they experience in the electron gun in order to get up to speed? For more on this subject, you might want to check our browser-interactive relativistic Accel-One problem solver at {http://newton.umsl.edu/~run/index.html} , and the theory pages attached.
The foregoing thoughts on lonely, fat, long and fast electrons are not really things I've had time to think much about, but they are interesting, and hence I would enjoy other perspectives on them, as well as suggestions for other "remarkable TEM facts" to add to the list!
Cheers. /philf :)
//\/\/\/\---} // Phil Fraundorf Physics & Astronomy/CME 314+5165044 philf-at-newton.umsl.edu \\ B503 U.Missouri-SL St.Louis MO 63121 USA http://newton.umsl.edu/~philf \\/\/\/\/\/\/\/---}
Greg -- I have been using a PC based image analysis package from Imaging Research Inc.(no relation)to do morphometry and automated gold particle counting on digitized EM negs for a couple of years. The use of digital image analysis (IA) vs. point counting is highly dependant on your images and what information you want to extract from them. If the structures of interest can be easily segmented from the rest of the image, then IA is the way to go. But if the measurement involves structures which need a human eye and brain (still the best image analysis system avaliable) to pick them out then point counting may be a much more efficient method. An example of the latter would be measuring lengths of plasma membrane sub-domains or the limiting membrane of non-descript organelles. While you can trace the membranes using the cursor of an IA system and get good data, its VERY tedious. You can do alot more sampling with point counting techniques. If the specimen was immuno-gold labeled, then IA would work well to count the gold particles along those membranes since the computer can recognize the particles quite well because they are very dense and have a very circular profile. But I would still count them manually if there weren't too many. There are IA systems out there that allow you to use point counting methods on digital images. Imaging Reserch has an add-on to their systems that does stereology, but I haven't had a chance to try it out yet. I suspect that the ability to segment out the structures of interest will still be crucial, at least for automated measurements. But as long as you can manually define "hits" of the sampling grid on the specimen these systems may be pretty useful. Sorry for being long-winded, I'm alone in a snow-bound lab.
Greg Martin Dept. Cell Biology and Anatomy Johns Hopkins School of Medicine gmartin-at-welchlink.welch.jhu.edu On Tue, 9 Jan 1996 Gargent-at-aol.com wrote:
} Are there any EM people heavy into Image processing and analysis. Currently } I am using a Kontron IBAS (version 2.0) and a KS400 Image analysis system. } Soon we will be incorporating a Telepathology system for communications } between the US, Switzerland, and Japan. Current Image imput for our routine } analysis come from EM micrographs, stored images, Kodak digital camera, from } the light microscope, and any source that allows us to capture, store, and } process images. We have several video camera, including a Sony 960 color } camera, B&W camera. We currently use PAL and NTSC formats. } } I would be interested in knowing how people use computerized Image Analysis } for Stereology and Morphometry. Would be interested in knowing how to get } around point counting techniques with Electron Micrographs. } } I would be willing to share IBAS macros with interested parties. } Gregory Argentieri } Sandoz Electron Microscopy lab } East Hanover, NJ 07936 } 201-503-8617 } Greg2NJ-at-AOL.COM } } } } Would welcome interested parties to exchange macros, ideas, image processing } tips etc. }
Received: from pkuns.PKU.EDU.CN by pccms.pku.edu.cn with SMTP id AA10706 (5.67b8/IDA-1.5 for zqliu); Sat, 13 Jan 1996 09:43:01 +0800 Received: from elaine12.Stanford.EDU by pkuns.PKU.EDU.CN with SMTP id AA11721 (5.67b8/IDA-1.5 for zqliu-at-pccms.pku.edu.cn); Sat, 13 Jan 1996 09:54:27 +0800 Received: (from wendyliu-at-localhost) by elaine12.Stanford.EDU (8.7.3/8.7.3) id SAA05523; Fri, 12 Jan 1996 18:12:05 -0800 (PST)
**** Very Important !!!! My Telphone and Fax Numbers have been changed recently !!!
Zhen Quan Liu, Director, Ph.D Tel (86) 10 275 1427 (Office) Physics Building Tel (86) 10 275 3727 (Home) Electron Microscope Laboratory Fax (86) 10 275 1615 (Office) Peking University Beijing 100871, China E-mail(Office): zqliu-at-pku.edu.cn Email (Home) wl-at-ibmstone.pku.edu.cn
If you haven't received many replies to your magnification problem, it may be because there are many sources of possible error in SEM magnification. What the mag circuit does is define the current through the scan coils and these establish a maximum scan ANGLE that raster will have as it leaves the final lens. How big an AREA this raster will cover on the specimen depends on how far the specimen is below the lens (the working distance, WD) and the square-root of the energy of the electrons in the beam.
Old SEMs where designed to only work at one working distance to eliminate the WD variable. Later it was found that one could get a measure of the WD (at a given beam energy and assuming no hysteresis in the magnetic circuit of the objective lens) by measuring the current in the final lens and later instruments used this information internally to compensate the scan current. Likewise the scan-currents were normalized to provide the same scan angles first at a few, and then later at a large variety of beam voltages.
To make all of this compensation work, there are a number of potentiometers in the scanning amplifier to allow the magnification to be adjusted in x and y under a number of standard conditions (certain Mags, WDs and kVs). It would seem at first glance that these pots may not be prperly adjusted on your instrument.
Normally SEMs are adjusted so that the magnification refers to the ratio of the length of a feature on a photograph (usually a Polaroid) produced by the microscopy divided by the length of the same feature in real life on the specimen. You have added the additional complication of a video printer. These can be useful but often only operate at video-scan-rate where SEM image distortion is high and where the printer makes specific assumptions aboutthe dimensions of the TV raster (number of lines, ration of H to V) that the SEM Manufacturer does not fulfill (and this is particularly true if you use a mixture of PAL (625 line) and NTSC (525 lines) equipment). You may also be using a computer-based image capture system that has a video-rate display which you print with your printer. On the assumption that this image-capture system is active (generates the scanning signals) rather than passive (listens to the scan signals generated by the microscope electronics) it seems likely that the misadjustments are in the scan-generation software of your image capture system rather than in the potentiometers noted above.
A lot of this information can be found a good basic SEM text.
Good LUck,
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
I realized that my first message about maxima and minima in SEM magnification might have been misleading, so I would like to provide some details. What we plot is the ratio of measured and nominal magnifications as a function of nominal magnification. The measured values come from black and white videoprints (Sony UP-850). The ratio ranges betwwen 0.75 and 0.94, depending on the microscope setting. The average and standard deviation of 73 ratios are 0.83 and 0.05, respectively. The curves exhibit the following behaviour : increase to a maximum about 200x, followed by a decrease to a mimium about 1000x, then an increase to about 3000x followed by a decrease down to 10000x. Please note that the resolution of the curve is rather low (8 data points in total), and it's not always easy to measure the magnification below 200x with a 10 micron spacing micrometer (which means the maximum at 200x is only observed when there are data points below 200x).
I hope this rings a bell to somebody...
Regards,
Antoine Ghanem ghanem-solvay-at-e-mail.com
Extra X400 information begins: Originator Name: Antoine (AGM) GHANEM Org Units: AC : NOH : LC-AN001 Organisation: NOHX400DEC Domain: BE/RTT/SOLVAY Node.Userid: IBMX400.184470
Message Id: 65239051106991/91343 MHS Importance: Normal Sent by Name: Antoine (AGM) GHANEM Org Units: AC : NOH : LC-AN001 Organisation: NOHX400DEC Domain: BE/RTT/SOLVAY Node.Userid: IBMX400.184470 Free Fmt Name: Antoine GHANEM Phone Number: 3422 Subject: SEM magnigication Recipients Name: INTERNET INTERNET Domain: GB/IBMX400/IBMMAIL Node.Userid: IBMMAIL.INTERNET Free Fmt Name: INTERNET INTERNET
I realized that my first message about maxima and minima in SEM magnification might have been misleading, so I would like to provide some details. What we plot is the ratio of measured and nominal magnifications as a function of nominal magnification. The measured values come from black and white videoprints (Sony UP-850). The ratio ranges betwwen 0.75 and 0.94, depending on the microscope setting. The average and standard deviation of 73 ratios are 0.83 and 0.05, respectively. The curves exhibit the following behaviour : increase to a maximum about 200x, followed by a decrease to a mimium about 1000x, then an increase to about 3000x followed by a decrease down to 10000x. Please note that the resolution of the curve is rather low (8 data points in total), and it's not always easy to measure the magnification below 200x with a 10 micron spacing micrometer (which means the maximum at 200x is only observed when there are data points below 200x).
I hope this rings a bell to somebody...
Regards,
Antoine Ghanem ghanem-solvay-at-e-mail.com
Extra X400 information begins: Originator Name: Antoine (AGM) GHANEM Org Units: AC : NOH : LC-AN001 Organisation: NOHX400DEC Domain: BE/RTT/SOLVAY Node.Userid: IBMX400.184470
Message Id: 50704151106991/92175 MHS Importance: Normal Sent by Name: Antoine (AGM) GHANEM Org Units: AC : NOH : LC-AN001 Organisation: NOHX400DEC Domain: BE/RTT/SOLVAY Node.Userid: IBMX400.184470 Free Fmt Name: Antoine GHANEM Phone Number: 3422 Subject: SEM magnification Recipients Name: INTERNET INTERNET Domain: GB/IBMX400/IBMMAIL Node.Userid: IBMMAIL.INTERNET Free Fmt Name: INTERNET INTERNET
Nancy - There is an error in the Web address listed - it should be ddsdx.uthscsa.edu, not ddxds.uthscsa.edu. It took me almost an hour to find it, then I realized what was wrong. Hope everyone else finds it ok -
BTW - I have tried to subsribe to the ,mail list for IT as listed on the Web page, but I keep getting error messages back - Any ideas?
Greg -- I have been using a PC based image analysis package from Imaging Research Inc.(no relation)to do morphometry and automated gold particle counting on digitized EM negs for a couple of years. The use of digital image analysis (IA) vs. point counting is highly dependant on your images and what information you want to extract from them. If the structures of interest can be easily segmented from the rest of the image, then IA is the way to go. But if the measurement involves structures which need a human eye and brain (still the best image analysis system avaliable) to pick them out then point counting may be a much more efficient method. An example of the latter would be measuring lengths of plasma membrane sub-domains or the limiting membrane of non-descript organelles. While you can trace the membranes using the cursor of an IA system and get good data, its VERY tedious. You can do alot more sampling with point counting techniques. If the specimen was immuno-gold labeled, then IA would work well to count the gold particles along those membranes since the computer can recognize the particles quite well because they are very dense and have a very circular profile. But I would still count them manually if there weren't too many. There are IA systems out there that allow you to use point counting methods on digital images. Imaging Reserch has an add-on to their systems that does stereology, but I haven't had a chance to try it out yet. I suspect that the ability to segment out the structures of interest will still be crucial, at least for automated measurements. But as long as you can manually define "hits" of the sampling grid on the specimen these systems may be pretty useful. Sorry for being long-winded, I'm alone in a snow-bound lab.
Greg Martin Dept. Cell Biology and Anatomy Johns Hopkins School of Medicine gmartin-at-welchlink.welch.jhu.edu On Tue, 9 Jan 1996 Gargent-at-aol.com wrote:
} Are there any EM people heavy into Image processing and analysis. Currently } I am using a Kontron IBAS (version 2.0) and a KS400 Image analysis system. } Soon we will be incorporating a Telepathology system for communications } between the US, Switzerland, and Japan. Current Image imput for our routine } analysis come from EM micrographs, stored images, Kodak digital camera, from } the light microscope, and any source that allows us to capture, store, and } process images. We have several video camera, including a Sony 960 color } camera, B&W camera. We currently use PAL and NTSC formats. } } I would be interested in knowing how people use computerized Image Analysis } for Stereology and Morphometry. Would be interested in knowing how to get } around point counting techniques with Electron Micrographs. } } I would be willing to share IBAS macros with interested parties. } Gregory Argentieri } Sandoz Electron Microscopy lab } East Hanover, NJ 07936 } 201-503-8617 } Greg2NJ-at-AOL.COM } } } } Would welcome interested parties to exchange macros, ideas, image processing } tips etc. }
Received: from pkuns.PKU.EDU.CN by pccms.pku.edu.cn with SMTP id AA10706 (5.67b8/IDA-1.5 for zqliu); Sat, 13 Jan 1996 09:43:01 +0800 Received: from elaine12.Stanford.EDU by pkuns.PKU.EDU.CN with SMTP id AA11721 (5.67b8/IDA-1.5 for zqliu-at-pccms.pku.edu.cn); Sat, 13 Jan 1996 09:54:27 +0800 Received: (from wendyliu-at-localhost) by elaine12.Stanford.EDU (8.7.3/8.7.3) id SAA05523; Fri, 12 Jan 1996 18:12:05 -0800 (PST)
**** Very Important !!!! My Telphone and Fax Numbers have been changed recently !!!
Zhen Quan Liu, Director, Ph.D Tel (86) 10 275 1427 (Office) Physics Building Tel (86) 10 275 3727 (Home) Electron Microscope Laboratory Fax (86) 10 275 1615 (Office) Peking University Beijing 100871, China E-mail(Office): zqliu-at-pku.edu.cn Email (Home) wl-at-ibmstone.pku.edu.cn
Microscopy & Microanalysis '96, the joint 54th Annual Meeting of the Microscopy Society of America, 30th of the Microbeam Analysis Society and the 23rd of the Microscopical Society of Canada / Societe de Microscopie du Canada, will be held August 11-15, 1996, in Minneapolis, Minnesota.
The deadline for receipt of papers (extended abstracts) is March 15, 1996. The Registration Bulletin / Call for Papers will be mailed automatically to members of the three Societies by the end of January.
For information contact:
Microscopy & Microanalysis '96 4 Barlows Landing Rd., Suite 8 Pocasset, MA 02644
I posed the following questions to the nih-image mailing list to clear up some of the current discussion. More info on the mailing list can be found at the bottom of the page. The response is from wayne Rasband, the author of NIH Image.
} Date: Fri, 12 Jan 1996 13:03:30 -0600 (CST) } Reply-To: nih-image-at-Soils.Umn.EDU } Originator: nih-image-at-soils.umn.edu } Sender: nih-image-at-Soils.Umn.EDU } Precedence: bulk } From: wayne-at-helix.nih.gov (Wayne Rasband) } To: Multiple recipients of list {nih-image-at-Soils.Umn.EDU} } Subject: Re: Import/processing questions } X-Comment: NIH Image Distribution List } Mime-Version: 1.0 } Status: O } } } Hey- } } } } On the microscopy list server (Microscopy-at-MSA.Microscopy.Com) we are } } currently having a discussion about Image's capabilities. They are as } } follows. } } } } 1. NIH Image supports 4, 8 and 16 bit TIFF (as well as PICT, pics, } } macpaint) images in both color and grayscale and 24 bit color images. Is } } that right? } } NIH Image can read 4, 8 and 16 bit grayscale TIFF files as well as 24 bit } color TIFF files. It can't read compressed TIFF fles. } } } has any one written macros for importing 30 bit (or more) color } } (or Greyscale) TIFF images? } } Not that I know of. Do such TIFF files exist? } } } I looked in the macros file on zippy, but none } } of them seem to do just this? } } } 2. Image processes 8 pixel neighborhoods surrounding the central pixel. } } The built-in filters process 3x3 (8 pixel) neighborhoods. The Convolve } command supports user defined kernels up to 63x63 (3968 pixel } neighborhoods). } } } } } 3. Image uses either 8 or 16 bit integers, depending on the input image, } } when doing image math, yes or no? } } The Image Math command uses real operations but scales the result to } 8-bits. V1.59 adds the ability to process 32-bit real images. } } } 4. UTHSCSA ImageTool for Windows sounds very much like Image, does it use } } NIH Image source code? } } I doubt it uses any nih-image source code since it's written in C++ } (nih-image is written in Pascal) and it doesn't look much like nih-image. } It does, however, appear to be in the spirit of nih-image. There is a link } to the Image Tool home page on the nih-image home page } (http://rsb.info.nih.gov/nih-image/). } } --wayne
Join the NIH-Image mailing list. by sending a message to listproc-at-soils.umn.edu with subscribe nih-image {first name} {last name} in the message body.
archived email (since 09/93) is available at the following:
Browse messages by date: {ftp://ftp.soils.umn.edu:pub/info/email-lists/nih-image/} {gopher://gopher.soils.umn.edu:10082/11/email-lists/nih-image}
-Kirk _____________________________________________ Kirk Rogers krogers-at-materials.ecn.purdue.edu OR kirk.a.rogers.1-at-purdue.edu Purdue University, School of Materials Science and Engineering, 1289 MSEE building, W. Lafayette, IN 47907-1289 OFFICE: 317-494-8751 FAX: 317-494-1204 http://materials.ecn.purdue.edu/~krogers
Hopefully the attached message from Brent Dove will clarify instructions for downloading Image Tool (IT), free image analysis software for Windows 95 and Windows NT....nkrs
} Return-path: {DOVE-at-uthscsa.edu} } Date: Fri, 12 Jan 1996 11:03:30 -0500 (CDT) } Date-warning: Date header was inserted by uthscsa.edu } From: "S. Brent Dove" {dove-at-uthscsa.edu} } Subject: IMAGETOOL } To: Nancy Smith {smithn-at-uthscsa.edu} } } Hello Nancy, } } I am being inundated with email. Please update your listserver with the following information. About how to get UTHSCSA ImageTool IT is available via anonymous ftp at=20 } } ftp://maxrad6.uthscsa.edu } } Please do not send email to dove-at-uthscsa.edu } } I have enclosed the original write up including the ftp address at the bottom. Sorry that was my fault for not including the ftp address. Thank you for getting the information to other microscopist. } } UTHSCSA } IMAGETOOL } } OVERVIEW } } UTHSCSA ImageTool (IT) is a free image processing and analysis program for Microsoft Windows 95=81 or Windows NT=81. IT can acquire, display, edit, analyze, process, compress, save and print gray scale and color images. IT can read and write over 22 common file formats including BMP, PCX, TIF, GIF and JPEG. Image analysis functions include dimensional (distance, angle, perimeter, area) and gray scale measurements (point, line and area histogram with statistics). ImageTool supports standard image processing functions such as contrast manipulation, sharpening, smoothing, edge detection, median filtering and spatial convolutions with user-defined convolution masks. IT also has built-in macro capabilities that allow the user to record repetitive tasks and playback saved macros to automate image analysis. } } ImageTool was designed with an open architecture that provides extensiblity via a variety of plug-ins. Support for image acquisition using either Adobe Photoshop plug-ins or Twain scanners is built-in. Custom analysis and processing plug-ins can be developed using the software development kit (SDK) provided (with source code). This approach makes it possible to solve almost any data acquisition or analysis problem with IT. } } ImageTool provides for geometric transformations such as rotate, flip vertical, flip horizontal and magnification up to four levels. All analysis and processing functions are available at any magnification factor. The program is a multiple document interface (MDI) application supporting any number of windows (images) simultaneously. =20 } } Spatial calibration is available to indicate real world dimensional measurements such as millimeters, microns, feet, miles, etc. for linear and area. Density or gray scale calibration can be done relative to radiation or optical density (OD) standards. } } IT version 1.1 now provides for object analysis and classification with over 20 morphological descriptors such as: area/perimeter, roundness, ferret diameter, compactness, major/minor axis length, centroid and many others. Any of these factors can be used automatically categorized and count objects within the image. } } ImageTool ver 1.1 supports the Data Translation DT3155 frame grabber for Windows NT. Other frame grabber board will be added in the coming months.= =20 } } ImageTool was written using Borland's C++ version 4.5 and the source code for the executable is available free of charge. IT was developed in the Department of Dental Diagnostic Science at The University of Texas Health Science Center, San Antonio, Texas. The program was developed by C. Donald Wilcox, S. Brent Dove, W. Doss McDavid and David B. Greer. } } UTHSCSA ImageTool is availabel via anonymous ftp at= ftp://maxrad6.uthscsa.edu } } S. Brent Dove Voice: (210) 567-3333 } Diagnostic Sciences Fax: (210) 567-3334 } University of Texas Email: dove-at-uthscsa.edu } Health Science Center Web: ddxds.uthscsa.edu } San Antonio, TX USA ftp: maxrad6.uthscsa.edu } }
Message-Id: {199601160327.UAA00999-at-ns1.indirect.com} Comments: Authenticated sender is {earosen-at-indirect.com}
Hi! Our graduate assistant had this floating around the school, not sure where he found it. I thought you might appreciate this considering you're working on your thesis.
150 THINGS (NOT) TO DO OR SAY AT OR FOR YOUR THESIS DEFENSE } 1) "Ladies and Gentlemen, please rise for the singing of our National Anthem..." 2) Charge 25 cents a cup for coffee. 3) "Charge the mound" when a professor beans you with a high fast question. 4) Describe parts of your thesis using interpretive dance. 5) "Musical accompaniment provided by..." 6) Stage your own death/suicide. 7) Lead the specators in a Wave. 8) Have a sing-a-long. 9) "You call THAT a question? How the hell did they make you a professor?" 10) "Ladies and Gentlemen, as I dim the lights, please hold hands and concentrate so that we may channel the spirit of Lord Kelvin..." 11) Have bodyguards outside the room to "discourage" certain professors from sitting in. 12) Puppet show. 13) Group prayer. 14) Animal sacrifice to the god of the Underworld. 15) Sell T-shirts to recoup the cost of copying, binding, etc. 16) "I'm sorry, I can't hear you - there's a banana in my ear!" 17) Imitate Groucho Marx. 18) Mime. 19) Hold a Tupperware party. 20) Have a bikini-clad model be in charge of changing the overheads. 21) "Everybody rhumba!!" 22) "And it would have worked if it weren't for those meddling kids..." 23) Charge a cover and check for ID. 24) "In protest of our government's systematic and brutal opression of minorities..." 25) "Anybody else as drunk as I am?" 26) Smoke machines, dramatic lighting, pyrotechnics... 27) Use a Super Soaker to point at people. 28) Surreptitioulsy fill the room with laughing gas. 29) Door prizes and a raffle. 30) "Please phrase your question in the form of an answer..." 31) "And now, a word from our sponsor..." 32) Present your entire talk in iambic pentameter. 33) Whine piteously, beg, cry... 34) Switch halfway through your talk to Pig Latin. Or Finnish Pig Latin. 35) The Emperor's New Slides ("only fools can't see the writing...") 36) Table dance (you or an exotic dancer). 37) Fashion show. 38) "Yo, a smooth shout out to my homies..." 39) "I'd like to thank the Academy..." 40) Minstrel show (blackface, etc.). 41) Previews, cartoons, and the Jimmy Fund. 42) Pass the collection basket. 43) Two-drink minimum. 44) Black tie only. 45) "Which reminds me of a story - A Black guy, a Chinese guy, and a Jew walked into a bar..." 46) Incite a revolt. 47) Hire the Goodyear Blimp to circle the building. 48) Release a flock of doves. 49) Defense by proxy. 50) "And now a reading from the Book of Mormon..." 51) Leave Jehovah's Witness pamphlets scattered about. 52) "There will be a short quiz after my presentation..." 53) "Professor Walcerz, will you marry me?" 54) Bring your pet boa. 55) Tell ghost stories. 56) Do a "show and tell". 57) Food fight. 58) Challenge a professor to a duel. Slapping him with a glove is optional. 59) Halftime show. 60) "Duck, duck, duck, duck... GOOSE!" 61) "OK - which one of you farted?" 62) Rimshot. 63) Sell those big foam "We're number #1" (sic) hands. 64) Pass out souvenir matchbooks. 65) 3-ring defense. 66) "Tag - you're it!" 67) Circulate a vicious rumor that the Dead will be opening, making sure that it gets on the radio stations, and escape during all the commotion. 68) Post signs: "Due to a computer error at the Registrar's Office, the original room is not available, and the defense has been relocated to (Made-up non-existent room number)" 69) Hang a pinata over the table and have a strolling mariachi band. 70) Make each professor remove an item of clothing for each question he asks. 71) Rent a billboard on the highway proclaiming "Thanks for passing me Professors X,Y, and Z" - BEFORE your defense happens. 72) Have a make-your-own-sundae table during the defense. 73) Make committee members wear silly hats. 74) Simulate your experiment with a virtual reality system for the spectators. 75) Do a soft-shoe routine. 76) Throw a masquerade defense, complete with bobbing for apples and pin-the-tail-on-the-donkey. 77) Use a Greek Chorus to highlight important points. 78) "The responsorial psalm can be found on page 124 of the thesis..." 79) Tap dance. 80) Vaudeville. 81) "I'm sorry Professor Smith, I didn't say 'SIMON SAYS any questions?'. You're out." 82) Flex and show off those massive pecs. 83) Dress in top hat and tails. 84) Hold a pre-defense pep rally, complete with cheerleaders, pep band, and a bonfire. 85) Detonate a small nuclear device in the room. Or threaten to. 86) Shadow puppets. 87) Show slides of your last vacation. 88) Put your overheads on a film strip. Designate a professor to be in charge of turning the strip when the tape recording beeps. 89) Same as #88, but instead of a tape recorder, go around the room making a different person read the pre-written text for each picture. 90) "OK, everybody - heads down on the desk until you show me you can behave." 91) Call your advisor "sweetie". 92) Have everyone pose for a group photo. 93) Instant replay. 94) Laugh maniacally. 95) Talk with your mouth full. 96) Start speaking in tongues. 97) Explode. 98) Implode. 99) Spontaneously combust. 100) Answer every question with a question. 101) Moon everyone in the room after you are done. 102) "Laugh, will you? Well, they laughed at Galileo, they laughed at Einstein..." 103) Hand out 3-D glasses. 104) "I'm rubber, you're glue..." 105) Go into labor (especially for men). 106) Give your entire speech in a "Marvin Martian" accent. 107) "I don't know - I didn't write this." 108) Before your defense, build trapdoors underneath all the seats. 109) Swing in through the window, yelling a la Tarzan. 110) Lock the department head and his secretary out of the defense room. And the coffee lounge, the department office, the copy room, and the mail room. Heck, lock them out of the building. And refuse to sell them stamps (NOTE: This is an inside gripe, based on conditions that existed in the ME department at WPI while we were there. Sorry.) 111) Roll credits at the end. Include a "key grip", and a "best boy". 112) Hang a disco ball in the center of the room. John Travolta pose optional. 113) Invite the homeless. 114) "I could answer that, but then I'd have to kill you" 115) Hide. 116) Get a friend to ask the first question. Draw a blank-loaded gun and "shoot" him. Have him make a great scene of dying (fake blood helps). Turn to the stunned audience and ask "any other wise-ass remarks?" 117) Same as #116, except use real bullets. 118) "Well, I saw it on the Internet, so I figured it might be a good idea..." 119) Wear clown makeup, a clown wig, clown shoes, and a clown nose. And nothing else. 120) Use the words "marginalized", "empowerment", and "patriarchy". 121) Play Thesis Mad Libs. 122) Try to use normal printed paper on the overhead projector. 123) Do your entire defense operatically. 124) Invite your parents. Especially if they are fond of fawning over you. ("We always knew he was such an intelligent child") 125) Flash "APPLAUSE" and "LAUGHTER" signs. 126) Mosh pit. 127) Have cheerleaders. ("Gimme an 'A'!!") 128) Bring Howard Cosell out of retirement to do color commentary. (NOTE: Because of recent events, this has to be changed to "Bring Howard Cosell back from the dead to do color commentary.") 129) "I say Hallelujah, brothers and sisters!" 130) Claim political asylum. 131) Traffic reports every 10 minutes on the 1's. 132) Introduce the "Eyewitness Thesis Team". Near the end of your talk, cut to Jim with sports and Alison with the weather. 133) Live radio and TV coverage. 134) Hang a sign that says "Thank you for not asking questions" 135) Bring a microphone. Point it at the questioner, talk-show style. 136) Use a TelePrompTer 137) "Take my wife - please!" 138) Refuse to answer questions unless they phrase the question as a limerick. 139) Have everyone bring wine glasses. When they clink the glasses with a spoon, you have to kiss your thesis. Or your advisor. 140) Offer a toast. 141) Firewalk. 142) Start giving your presentation 15 minutes early. 143) Play drinking thesis games. Drink for each overhead. Drink for each question. Chug for each awkward pause. This goes for the audience as well. 144) Swoop in with a cape and tights, Superman style. 145) "By the power of Greyskull..." 146) Use any past or present Saturday Night Live catchphrase. Not. 147) Stand on the table. 148) Sell commercial time for your talk and ad space on your overheads. 149) Hold a raffle. 150) "You think this defense was bad? Let me read this list to show you what I COULD have done..." } (FINAL NOTE: Depending on the subject of your thesis, some of these things, such as tap dance, virtual reality, or reading from the Book of Mormon might be entirely appropriate, of course.) }
} On 15 Jan 1996 Hans Brinkies wrote: } } } Is there any evidence available that the extensive use of TEM's } } and/or SEM's may lead to the development of cataracts ? } } } } } } Hans Brinkies } } This is an interesting point. Some time ago someone asked a similar } question but I cannot remember seeing any discussion on the issue. Does } anyone know about the long term health effects of EM usage? Have any studies } been done on any aspect of this (eyesight, radiation exposure, anything else } I haven't thought of)? } } Ian MacLaren,
EMSA (now MSA) published a "Handbook of X-ray safety for Electron Microscopists" by D.F} Parsons, V.A. Phillips, and J.S. Lally in 1973. Its a short pamphlet and I don't know if it is still available. A few points from the handbook:
In regards to the need to perform an annual survey for x-ray leaks: "It should be noted that about 50% of those monitoring their microscopes found detectable leaks once or more". They reccomend film badges but I have never been at an institution that did this for their EM people.
Under the Health Checks chapter: "An annual eye exam is desirable. Vision defects should be recorded and the lenses checked for opacities (cataracts)". No references or data supporting this statement are given.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
Dear Phil, } } Lonely electrons: [snip] } Under some illumination conditions there may be no more than 1 beam electron } in the column at a time!
With our HVEM (1.2 MV) and its ~3m column this turns out to be cor- rect--especially since we use low-dose conditions for ED work.
} Hence diffraction patterns in the TEM are } basically formed by individual electrons interfering with themselves!
But wait; there's more! The electrons are boiled off our W filament as particles, interfere with themselves as waves when scattering off the crystal, then strike a film grain as particles. ED is the quintessential demonstration of the particle-wave duality.
} Fat electrons: The transverse coherence widths of electrons which make } possible electron phase contrast (HREM) lattice imaging and probably } electron holography might also be seen as lateral broadening of individual } electron wave-packets via the uncertainty principle, which results because } we know too much about their transverse momentum! [snip]
What are the predictions for hollow-cone illumination where we know the magnitude of the transverse momentum, but not its direction?
} [snip] } Fast electrons: a back of the envelope calculation for 300 keV electrons } gives gamma = (300+511)/511 = 1.587, so that they travel at w = c } Sqr[1-(1/gamma)^2] = 0.777 c or [lightyears per inertial year] of elapsed } time.
Of course, for 1.2 MeV electrons the speed is even closer to c. Yours, Bill Tivol
Sorry for bugging once again all those helpful people who have given advice on the conversion of image file formats. I specifically have a question about AUTOCAD drawings. If anyone knows how to convert these to more common image files, please get in touch with me directly. Thanks, Alfred
Alfred Kracher akracher-at-iastate.edu http://www.public.iastate.edu/~akracher
Yes, the ribbons have changed. If you complain enough and to the right person at Kodak, Kodak will send you a disk with new color look uop tables that can be downloaded to the printer. Your b/w prints will come out with a slight blue tint instead of the green tint, but the blue tint is less objectionable. -Michael Cammer {null signature file}
On Thu, 11 Jan 1996, Jon Krupp wrote:
} Hi: } } Has anyone run into a problem printing grayscale images on a Kodak/Codonics } printer? } } Ours have started looking tinged with green/yellow right out of the } printer. I have read that the formulation of the ribbons may have changed } and that the result is this sick green color. } } I would like to avoid swapping color/grayscale ribbons if possible but my } users don't like the color cast to their pictures. Any ideas for a } solution? } } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
Yes, the ribbons have changed. If you complain enough and to the right person at Kodak, Kodak will send you a disk with new color look uop tables that can be downloaded to the printer. Your b/w prints will come out with a slight blue tint instead of the green tint, but the blue tint is less objectionable. -Michael Cammer {null signature file}
On Thu, 11 Jan 1996, Jon Krupp wrote:
} Hi: } } Has anyone run into a problem printing grayscale images on a Kodak/Codonics } printer? } } Ours have started looking tinged with green/yellow right out of the } printer. I have read that the formulation of the ribbons may have changed } and that the result is this sick green color. } } I would like to avoid swapping color/grayscale ribbons if possible but my } users don't like the color cast to their pictures. Any ideas for a } solution? } } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
Does anyone have any experience with Technovit resin? Someone here has heard about it, but we don't have any "real" info from users. They want to use it for LM immuno...enzyme-based, as the tissue is massively autofluorescent. Thanks for the help! Tamara Howard
The Department of Biological Sciences is seeking applications for Director of the Electron Microscope (EM) Facility as an academic staff appointment. Broad experience with a diversity of biological systems and EM techniques and Ph.D. in the Biological Sciences required, postdoctoral experience preferred. Responsibilities include: 1) management of the Biological Science EM facility, 2) research support for and collaboration with university faculty, 3) instruction of faculty and students in the use of EM. An independent funded research program is desirable. The Biological Sciences Electron Microscopy Facility consists of two laboratories totaling 2000+ square feet, SEM and TEM scopes and support equipment.
Applicants should submit a curriculum vitae, statement of professional interests, and three letters of reference to:
Chairperson, EM Director Search Committee Department of Biological Sciences University of Wisconsin--Milwaukee P.O. Box 413, Milwaukee, WI 53201.
The deadline for receipt of applications is February 16,1996.
Additional information about the Biological Sciences Department is available at http:\www.uwm.edu\Dept\Biology\. The University of Wisconsin-Milwaukee is an Equal Opportunity/Affirmative Action Employer. Women and minority scientists are encouraged to apply for these positions. The names of those nominees and applicants who have notrequested that their identities be withheld and the names of finalists will be released upon request.
The Department of Biological Sciences is seeking applications for Director of the Electron Microscope (EM) Facility as an academic staff appointment. Broad experience with a diversity of biological systems and EM techniques and Ph.D. in the Biological Sciences required, postdoctoral experience preferred. Responsibilities include: 1) management of the Biological Science EM facility, 2) research support for and collaboration with university faculty, 3) instruction of faculty and students in the use of EM. An independent funded research program is desirable. The Biological Sciences Electron Microscopy Facility consists of two laboratories totaling 2000+ square feet, SEM and TEM scopes and support equipment.
Applicants should submit a curriculum vitae, statement of professional interests, and three letters of reference to:
Chairperson, EM Director Search Committee Department of Biological Sciences University of Wisconsin--Milwaukee P.O. Box 413, Milwaukee, WI 53201.
The deadline for receipt of applications is February 16,1996.
Additional information about the Biological Sciences Department is available at http:\www.uwm.edu\Dept\Biology\. The University of Wisconsin-Milwaukee is an Equal Opportunity/Affirmative Action Employer. Women and minority scientists are encouraged to apply for these positions. The names of those nominees and applicants who have notrequested that their identities be withheld and the names of finalists will be released upon request.
MSA published an updated verion of the Safety Handbook edited by Joe Mascorro and Vernon Barber in 1994. It is available from San Francisco Press, Inc. Box 426800, CA 94142-6800. I believe it costs $17.00.
In our central facility, all of our electron microscopes are evaluated by radiological professionals annually (as mandated by state law) who use extremely sensitive detection devices. They have never detected x-rays above background levels. Nonetheless, best to be safe and cautious.
} } On 15 Jan 1996 Hans Brinkies wrote: } } } } } Is there any evidence available that the extensive use of TEM's } } } and/or SEM's may lead to the development of cataracts ? } } } } } } } } } Hans Brinkies } } } } This is an interesting point. Some time ago someone asked a similar } } question but I cannot remember seeing any discussion on the issue. Does } } anyone know about the long term health effects of EM usage? Have any studies } } been done on any aspect of this (eyesight, radiation exposure, anything else } } I haven't thought of)? } } } } Ian MacLaren, } } } EMSA (now MSA) published a "Handbook of X-ray safety for Electron } Microscopists" by D.F} Parsons, V.A. Phillips, and J.S. Lally in 1973. Its } a short pamphlet and I don't know if it is still available. A few points } from the handbook: } } In regards to the need to perform an annual survey for x-ray leaks: "It } should be noted that about 50% of those monitoring their microscopes found } detectable leaks once or more". They reccomend film badges but I have } never been at an institution that did this for their EM people. } } Under the Health Checks chapter: "An annual eye exam is desirable. Vision } defects should be recorded and the lenses checked for opacities } (cataracts)". No references or data supporting this statement are given.
############################################################################# John J. Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
JOB DESCRIPTION: Sample preparation of semiconductor materials and devices, both plan view and cross-section, for TEM. Develop improved methods of sample preparation. TEM operation to facilitate this development will be encouraged. Photographic processing and filing of TEM negatives and prints. Maintenance of sample preparation and dark room equipment and supplies
JOB REQUIREMENTS: 2+ years experience in TEM sample preparation and photographic processing required. Familiarity with tripod, dimpling and FIB techniques desired. This position requires strong organizational skills, and in particular, the ability to work on multiple tasks while maintaining precise records and tracking procedure. Experience in microelectronics industry a plus.
Associate degree in physics, chemistry or related subject.
We are considering establishing an image archiving proceedure based on CD's. In addition the CD burner would be used by an electronic publishing class for prototype CD's.
The choices in hardware seem to have increased a lot recently and prices have really fallen.
If you have any sage advice concerning general guidelines or specific choices, I would like to hear from you.
Thanks,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 jmkrupp-at-cats.ucsc.edu
Message-Id: {199601161813.LAA18691-at-ns2.indirect.com} Comments: Authenticated sender is {earosen-at-indirect.com}
To the list Here is something else for you to peruse through if you are not busy and are interested in a laugh
How do you like this?? A friend mailed it to me. Its long.
FORREST in EVERYONE'S LIFE
Forrest Gump Life is like a Box of chocolates... Forrest Dahmer People are like a box of chocolate, YUM! Forrest Simpson Mmmmm, choolate Forrest the Hun Chocolate all mine! Forrest Simmons Chocolate is bad!, EXERCISE EXERCISE! Forrest Rivera People who like Chocolate..Next on 'Forrest' Forrest Jackson Little kids like my box of chocolates Forrest Hefner Keep the chocolate, lose the box. Forrest Shakespeare Chocolate, or no chocolate that's the question Forrest Of Borg All chocolates must be assimilated Forrest Presley Hunk a hunk of milk chocolate Forrest Zen I am one with the chocolate Forrest McClain I used to be a box of Chocolates Forrest Ventura Chocolates..Alll-Riighty then... Forrest Lauper People just wanna have chocolate Forrest Turner What's chocolate gotta do, gotta do with it? DR.Forrest McCoy Dammit jim, I'm a Dr., not a box of chocolate Forrest Spock Logically speaking, we are all chocolate Forrest Scotty The box, she's breaking apart Capt'n Forrest Christ Let he without sin, eat the first chocolate Forrest Rooney Why is it, that we are all chocolates? Forrest Butler Frankly Scarlett, I don't like chocolate Forrest O'Hara Tommorrow, is another box of chocolates. Forrest Lee Fight with your inner chocolate Forrest Clinton I didn't inhale the cream centers Forrest Doo Roinks Raggy, Rocolates! Forrest Pig Life is a box of chok-choa-che..candy Forrest Marx That's the weirdest box of chocolates I've ever seen.... Forrest Nicholson You want chocolate, you can't handle chocolate Forrest Copperfield Poof, the chocolates are gone! Forrest X We didn't land in the box of chocolate The box of chocolate landed on us! Forrest Hitler White Chocolates only! Forrest the Frog Someday we'll find it, The chocolate connections The plain ones, The cream filled....and me... Forrest Eastwood I know what your thinking.. Did he eat five chocolates, or did he eat six Well let me ask you... Do you feel hungry PUNK?..well...DO YOU? Forrest Barney I'm cream filled, your with nuts. We're a box of chocoluts Forrest Adam and Eve ADAM=Chocolates are forbidden EVE=Just eat one.... Forrest Moses I command the chocolates to seperate! Forrest Noah 2 creams, 2 nuts, 2 coconuts, 2 peanut butter Forrest Ali I am the chocolate boxer! Forrest on phonics Lief es lyk a boks uv chakolets Forrest PsychicLine Yes, I knew you were a chocolate 1-900-FORREST oooh, can I suck your cream filled chocolates? Forrest DatingGame Bacholer number two... if I was a piece of chocolate.. What would you fill me with? Forrest Alimony The Box is mine! Forrest Adultry You just can't have just one chocolate. The Forrest plague Ewww..these Chocolates are bad Chief Justice Forrest Thomas I never touched her milk-duds! Forrest Andrews The Hills are alive..like a box of chocolates Forrest Allen Chocolate, huroof.. Forrest Costello Whose eating chocolates? Forrest Abbot No, who is not eating chocolate Forrest Vader Luke, I am your chocolate Forrest Yoda There is a dark chocolate, and a light chocolate.. Forrest Butthead Uh...life is like a box of um..chocolates Forrest Beavis Heh heh, you said Box Forrest Frued Is life really a box of chocolate.. or is it your mother you want? Forrest SkeeLo I wish I had a box of Chocolates Forrest Trebeck The answer:This is like a box of Chocolate The Question:What is Life... Forrest Hillary Hey Bill...those are my Chocolates! Forrest Fudd Wife is wike a box uv chocowates Forrest Calvin It's not a box of chocolates, It's a transmorfgorizing ray!! Forrest Tannen Chocolates...McFly Dr. Forrest Ruth Chocolate is nice, but sex is better! The Forrest Zone There is another dimension, beyond that which is known to man, It's a dimension of cream-filled bon-bons, Or nutty carmel turtles, and it lies in the white cardboard box, in the pit of my lap. It is a candy coated center of comparision, And it is a place we call, The Forrest Zone. Forrest Latin ifela sia a oxba foa hocolatesa Forrest Lincoln Forescore and several chocolates ago, Forrest Satan Life is a box of melted chocolates (Sign) Forrest Churchlady Chocolates...well, isn't that special Forrest Wayne Sh-yeah..and life is like a box of chocolates as if... Forrest Garth Hey mister chocolate man..whose trying to kill you?...I don't know but her better not... Forrest Bond Lifes a box of exploding chocolate Forrest Smurf Smurf is a smurf of smurfs Forrest '95 The box is the same, The chocolates are upgraded.... Forrest Lennon Imagine there's no chocolate. Forrest Hall Will you take the Box of Chocolates... or what's behind CURTAIN NUMBER TWO? Forrest Press your Luck Box of Chocolates! No Whammies...STOP! Forrest of Fortune LIFE I_ LIKE _ __X _F CH_C_L_TE_ Forrest Popeye I yam a box of Chocolets...eg eg eg eg Forrest Ice If you got a chocolate, Yo I'll box it. Check out my life, As my D.J Rocks it! Forrest Ross Life is a happy little box of chocolates Forrest Bill and Ted Dude, Life's totally a box of Chocolate! ---EXCELLENT!---- ___AIR GUITARS TRIUMPHANTLY_____ Forrest Fife This Box of Chocolates is a lethal weapons! Forrest Stooges Look Chocolates....nyuk nyuk nyuk Scram Wise guy **BOink** Leave him ALone Moe! Oh you want in on it too...**SLAP**/CRASH/ Forrest Reagan Life is a box of um..... Forrest Bush This will not be another Box of Chocolates! Forrest Limbaugh Life may be a box of Chocolates... But the Democrats are all nuts... Forrest COPS on location Bon bons-bon bons whatcha going to do, whatcha gonna do, when someone eats you! Forrest Fued Life is like a? Box of Chocolates... Oh Good ANswer..Good Answer! Box of chocolates...SURVEY SAYS? XX|BOX OF CHOCOLATES|XX=} 54 Forrest Native Americans These chocolates are moving to a smaller box.
It depends on what you have and what you want. If you have a flat line drawing in AutoCAD and just want to get it in a form you can print out, then you just need to choose Export from the File menu (in R13 for Windows) and export it as an EPS (encapsulated Postscript) file. This will print directly on most Postscript printers. You can also save it as a BMP file.
If you have a flat line drawing in a DXF file, then you will need to convert it from a geometry file (which is what DXF is) to an image. You can do this by loading it into AutoCAD and then exporting it as above, you can use a file conversion program such as HiJaak Pro, or you can use one of the DXF utilities at the Viewpoint/Avalon site I mentioned before.
If you are in AutoCAD, have a 3D object, and want to save a rendering, then you have a couple more options. You can render in AutoCAD, either with the native renderer or with RenderMan (I can't remember whether or not R13 comes with RenderMan bundled or not), and save it as a BMP or EPS. You can export the geometry file into another geometry format, load that file into a different renderer, render there, and save the file as an image. I, for instance, do much of my modeling in AutoCAD, and render using 3D Studio or Lightwave 3D. There are a number of fairly good public domain renderers as well; the best known among the folk I travel with is POV.
If you have a geometry saved as a DXF file and want a rendered image, remember that DXF is a *geometry* not *image* file. While, for line drawings, many convertors will just do a simple projection (which is how HiJaak Pro and others work), if you really want an image with shaded graphics, you will have to make a rendering and save the image as an image from your renderer.
On Tue, 16 Jan 1996, Alfred Kracher wrote:
} Sorry for bugging once again all those helpful people who have given advice } on the conversion of image file formats. I specifically have a question } about AUTOCAD drawings. If anyone knows how to convert these to more common } image files, please get in touch with me directly. } Thanks, Alfred } } Alfred Kracher } akracher-at-iastate.edu } http://www.public.iastate.edu/~akracher } } }
Has anybody using Ralph knives to cut resin sections, used ultramicrotome -quality glass for making their knives? A sales person has tried to convince me that using this quality of glass 'should' provide good knives as opposed to 'histology - grade' glass. I pose this dilemma as I have found that 'ultramicrotome - grade' glass is locally cheaper. Does anyone on the list have an opinion and/or advice as to a good source of reliable glass for the Ralph knives. Regards,
Brett- I guess I qualify as a salesperson, since we manufacture Ralph Knife Makers, and also sell glass for them. However, I am a professional manager, not a salesman, and I am interested primarily in selling equipment, not consumables, and it is important to me that the equipment we sell works properly. With that "introduction," let me give you my opinion. "Ultramicrotome glass" and "histology glass" are labels that any vendor can put on any glass- that is, there are no rules governing how these terms are applied. According to conventional wisdom, "ultramicrotome glass" should be the highest quality, since cryoultramicrotomes are much more sensitive to glass quality than are the rotary microtomes used in histology. In our catalog, the ultramicrotome glass is of higher quality than the histology glass, although both begin life as "float glass" and then are cut for use in microscopy. That all being said, there can be tremendous quality differences between glass obtained from different sources.If you are concerened, I would recommend that you ask your salesperson for a sample piece of glass. Steven Slap, Vice-President Energy Beam Sciences, Inc.
Message-Id: {199601171524.JAA19206-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 01:36 PM 1/16/96 +0000, you wrote: } Disregard the forrest gump humor I was supposed to send it to a } single person and not to the whole group} } } } } Cheers ;o) :o) %o) } Eric } ************************ Oh, sure we're gonna disregard it now!
Hello, I was wondering if anyone is using a Macintosh-based software for image aquisition and manipulation that they are happy with? I have been using Metamorph (Love it!) but now need something for the Mac. Thanks in advance.
Ruth Hughes Central Institute for the Deaf 818 S. Euclid St. Louis, MO 63110 e-mail RMH-at-CIDMV1.WUSTL.EDU
Does anyone have a source for amorphous Ge grids at { { $100's? If not, does anyone know a good source for Ge--not necessarily the ultra- pure stuff? TIA. Yours, Bill Tivol
A veterinarian friend of mine has a son who wants a good stereomicroscope for examining insects. Any offers would be appreciated under $200, especially if it is a zoom model.
Dear Thomas, } } EMSA (now MSA) published a "Handbook of X-ray safety for Electron } Microscopists" by D.F} Parsons, V.A. Phillips, and J.S. Lally in 1973. Its } a short pamphlet and I don't know if it is still available.
It was still available about a year ago from San Francisco Press.
} A few points } from the handbook: } } In regards to the need to perform an annual survey for x-ray leaks: "It } should be noted that about 50% of those monitoring their microscopes found } detectable leaks once or more". They reccomend film badges but I have } never been at an institution that did this for their EM people.
We wear them routinely at the HVEM, and our users carry dosimeters on themselves in the scope room. We have had only one incident where } ~1mr was measured, and that was apparently not from the scope. Yours, Bill Tivol
The amount of radiation exposure from Probes, SEMs and all, is minimum, but it still has to be monitored by certified individuals. Most states consider the equipment as minimumal threat devices and do not require any testing...
You must be careful, in some states like Texas and Colorado, you must be a RSO (radiation safety officer) for that specific piece of equipment (i.e. Cameca, JEOL, ARL, ISI, .....) to do a survey or monitor the equipment. You must have a minimum of 5 years in repair and calibration and take a course in radiation safety and in certain cases you must take a test.
Some other states like California, they would like you to also have Hazwoper/Hazmat certification also, but is not required by law. It is incase their is a lawsuit. You could be taken to the cleaners without it.
After coming back to work and seeing this on my email, it was a welcome treat. There were bats in the belfry from staying in the house after all the snow we had here in Baltimore. Yes, I do know there are people out there who will get more snow in one month than we usually get all winter. So, thanks for your mistake.
We are thinking about purchasing a Dage DSP-2000 processor for image processing of light microscopic video images. Several features seem useful for our images: averaging (particularly useful), integration, and gating. It's about $6,000, and I'd like to look at similar signal averagers for video before ordering the Dage product. Any suggestions from readers of this list? Thanks for your help.
-- Nancy L Desmond, Ph.D. nld-at-virginia.edu Department of Neurosurgery 804.924.5607 (voice) 804.982.3829 (fax) University of Virginia Health Sciences Center, Box 420 Charlottesville, VA 22908
I'm just about to put a work study student on a project to digitize our standards' library. Once digitized, my libray might then be put on the web (with other libraries ???? ) But, before I do I can well imagine at least a hundred different formats for the database entry. Has this been done before?? Can anyone suggest the most useful format for searching the database??
cheers, shaf {\/} /\ {\/} /\ {\/} /\ {\/} cognito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
To those of you who are replying to people complaining about a post. PLEASE PLEASE try and remember to post the reply to the sender and not to the list. Some mailers are dumb and assume the list IS the sender, but others are not. Please check the configuration of your email software.
Many thanks.
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu, John.F.Mansfield-at-umich.edu or jfmjfm-at-umich.edu they all reach me! URL: http://wwwpersonal.engin.umich.edu/~jfmjfm/jfmjfm.html
The best place to start for Macintosh imaging is NIH Image. It is public domain software from NIH and is available by anonymous FTP from:
zippy.nimh.nih.gov
There is a WWW home page, as well:
http://rsb.info.nih.gov/nih-image/
It can inherently acquire images via frame grabbers (NTSC,PAL) as well as a host of other video-type inputs if you have the appropriate Photoshop plug- in modules (PIM). There are also SEM/EDS interfaces (4pi analysis) for digital acquistion using PIMs. It is able to open TIFF and PICT files directly, any 8- or 16-bit bitmap file and "many" other formats through import PIMs. A wide range of spatial and intensity information extraction is available as well as image processing functions. The software is inherently very useful and becomes extremely useful with the built-in Pascal-like macro language, PIMs and source level programming (the Pascal source code is public domain, as well). The manual includes a fairly comprehensive list of commercial imaging products as well. Best of all, getting up and running is very straightforward, particularly for someone who has already been using imaging software. Even better than best of all is the worldwide support network available through the NIH Image listserver.
My personal rule-of-thumb commercial value of the software would be in the $1000 to $2000 price range, based on the feature set of NIH Image and the comparable feature set for commercial software (Mac,Wintel, etc).
Have fun.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
I received this job description from someone who is not on the net. If any one is interested all the pertinent facts are listed below.
Position description
A Research Assistant position is available in a research laboratory in the Gerontology Division at the VA Medical Center, Palo Alto, CA. Studies carried out in this laboratory focus on cell biology questions relating to lipid pathways in cells and on age-related changes in cells. The position will be available in April or May of 1996, and some overlap with the retiring research assistant is desired. The position requires someone with well grounded electron microscope (EM) skills with experience and an interest in carrying out related morphological techniques including immunocytochemistry (at the light microscope level and EM level using cryoultramicrotomy), fluorescence microscopy, and in situ hybridization methodology. Persons applying for this job must also have basic biochemical skills, be adept at problem solving, flexible in working on several projects simultaneously, and willing to learn new techniques and use new equipment when necessary.
Employment will be through the Palo Alto Institute for Research and Education (PRAIRIE). Salary and hours are negotiable and dependent on training and experience. Please send resumes to Eve Reaven, PhD, VA Medical Center, (GRECC, 182B), 3801 Miranda Ave., Palo Alto, CA 94304. (FAX = (415) 855-9437; telephone = (415) 493-5000 x64144)
Mei Lie Wong Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 email wong-at-msg.ucsf.edu
If you have questions about the opening described below, contact me by e-mail, but all application letters and resumes should be sent as hardcopy to be considered. Note that the position involves physical sciences and that we do not intend to consider applicants with a biological sciences background.
--------------------------------------
TRANSMISSION ELECTRON MICROSCOPY
THE PENNSYLVANIA STATE UNIVERSITY
The Materials Characterization Laboratory at Penn State is seeking candidates for a staff position in transmission electron microscopy. The successful applicant will take direct responsibility for use and support of the laboratory's 200 kV field-emission instrument, which is equipped with EDS and PEELS detectors and a SS-CCD camera for digital acquisition. The person selected will be expected to work with users from a variety of disciplines on a wide range of materials problems. In addition, candidates should be prepared to give advice on and assistance with sample preparation, technique selection, and analysis of images, diffraction patterns, and spectra. The person hired will also be able to pursue collaborative or independent research as other duties allow.
Requirements include a B.S. and preferably an M.S. in Materials Science or a related field, and additional experience beyond the degree is preferred. The position is not intended to be a postdoctoral appointment, although this may be considered if other suitable candidates are not found. Candidates should have extensive experience with transmission electron microscopy and must be skilled in one or more of the following areas: high-resolution imaging and simulation, quantitative EDS, PEELS, electron holography, field-emission TEM operation.
The position will be filled initially on a two-year appointment, but it is intended that the position will be made permanent if performance is satisfactory. Respond to Prof. A. H. Carim, 118 Steidle Building, The Pennsylvania State University, University Park, PA, 16802.
------------------------------------------------------------------------ Prof. Altaf H. Carim tel.: (814) 863-4296 Dept. of Materials Science & Engineering fax: (814) 865-0016 118 Steidle Building e-mail: carim-at-ems.psu.edu The Pennsylvania State University University Park, PA 16802 ------------------------------------------------------------------------
Dear All, As I understand it, the concern over cataracts in an EM lab was from the UV radiation from the evaporator, not X-rays from the EMs. The Safety in the EM Handbook has a discussion of various eye hazards from lasers and UV sources, but we always use a pair of welders goggles when evaporating carbon, since the arc is dangerous to the eyes, as is a welder's arc. A few years ago I tested my sputter coater for UV light while it was operating and was dismayed at how much UV was coming out of the thick glass. I advise people not to stare intently at their samples while they are coating.
A few months ago several people on the Listserver tried to see if their physical ailments were common to other microscopists, problems such as shoulder joint and neck pains. There were many different ailments, but no common threads. We're all just getting older, I guess. Hope this helps, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Yes. An excellent package for color image acquisition and analysis for the Macintosh is made by Signal Analytics. I can send you a demo disk and literature if you are interested. I represent Signal as well as several other software companies.
John Libert
At 11:19 AM 1/17/96 -0500, Ruth Hughes wrote: } Hello, } I was wondering if anyone is using a Macintosh-based software for } image aquisition and manipulation that they are happy with? I have been } using Metamorph (Love it!) but now need something for the Mac. } Thanks in advance. } } Ruth Hughes } Central Institute for the Deaf } 818 S. Euclid } St. Louis, MO 63110 } e-mail RMH-at-CIDMV1.WUSTL.EDU } } }
The old email address ( go4pi-at-applelink.apple.com) is no longer valid.
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
If you are willing to use a computer as part of your video averaging scheme, there are a number of boards which will do video-rate signal averaging (e.g., Scion 301-695-7870, D1887-at-applelink.apple.com for Mac and probably Wintel "soon"; Matrox 800-804-6243, imaginginfo-at-matrox.com for Wintel). You could also consider on-chip integration, but that is camera specific.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Regarding the thread about radiation safety and electron microscopes:
Don't assume that your state doesn't require some type of registration and periodic inspection of your electron microscope for radiation safety. It could cost you a hefty fine from the government agency in your state responsible for such matters if you don't register your instruments and have a program in place to assure compliance with the regulations.
The instrument manufacturers generally don't mention anything about the requirements to register their instruments when you purchase and install them. It's your responsibility to find out what your state requires.
Hope this saves somebody some headaches and some money.
Jim Stets Air Products and Chemicals, Inc. Allentown, PA stetsjr-at-ttown.apci.com
By measuring the intensity distribution around a Debye - ring over a range of goniometer tilt positions I can produce a partial pole figure of the selected specimen area. The diffracted intensity in a certain direction is taken as the difference between the reading at the ring and reading just inside it. The readings can be accurate to about 2 (azimuth angle).
Now, as the specimen is tilted through a decided range it becomes necessary to make some corrections to the measured intensities since the specimen thickness increase, as do the illuminated volume. Absorption and extinction changes should be taken into account.
It is possible to estimate such a correction by some simple experiments. However, what I would like to know is if there exist any software that can calculate the needed corrections. If so, is this calculation/software available free of charge?
Yours sincerely
P. Baggethun Manchester Materials Science Centre UMIST
Message-ID: {MAPI.Id.0016.00683536202020204437333230303030-at-MAPI.to.RFC822} Read-Receipt-To: Jean-Louis Beek {ah56-at-solo.pipex.co.za} Priority: Normal To: microscopy-at-Sparc5.Microscopy.Com MIME-Version: 1.0
Dear Bill,
You replied to:
[snip] } } Under some illumination conditions there may be no more than 1 beam electron } } in the column at a time! } With our HVEM (1.2 MV) and its ~3m column this turns out to be cor- } rect--especially since we use low-dose conditions for ED work. Wow! The current of electrons hitting the specimen must be quite low. How do you estimate it?
[snip] } What are the predictions for hollow-cone illumination where we know } the magnitude of the transverse momentum, but not its direction? I haven't thought about it yet, but I think hollow-cone illumination was being used by Dr. Murray Gibson's group at U. of I. (Champaign-Urbana) to vary the coherence widths (and lengths?) of their electrons. Check with them.
I should add for listserver readers as well, that I am interested in developing these informal observations on fast, lonely, fat and long electrons a bit further, and of course to add new ones as well. Feedback is invited! Some of you may see a bit more of this on paper in days ahead, but in the meantime we're sponsoring a (hopefully ever-improving) version of the list through our "relativity rap" web page at {http://newton.umsl.edu/~run/rap.html} .
Cheers. /philf :)
//\/\/\/\---} // Phil Fraundorf Physics & Astronomy/CME (314)5165044 philf-at-newton.umsl.edu \\ B503 U.Missouri-SL St.Louis MO 63121 USA http://newton.umsl.edu/~philf \\/\/\/\/\/\/\/---}
} we make diamond grids. We could make amorphous Ge grids if you want. } What do you want them for? Is there a general demand for these grids? } Dear Richard, I want the grid for measuring spherical aberration from the zeros of the contrast transfer function by Krivanek's method. I saw this described in Ultramicroscopy 38 (1991) 225-233--which was more easily obtained than Krivanek's paper in Optik 45 (1976) 97ff. There may be some general demand for these grids, but I cannot tell you how much. Yours, Bill Tivol
By measuring the intensity distribution around a Debye - ring over a range of goniometer tilt positions I can produce a partial pole figure of the selected specimen area. The diffracted intensity in a certain direction is taken as the difference between the reading at the ring and reading just inside it. The readings can be accurate to about 2 (azimuth angle).
Now, as the specimen is tilted through a decided range it becomes necessary to make some corrections to the measured intensities since the specimen thickness increase, as do the illuminated volume. Absorption and extinction changes should be taken into account.
It is possible to estimate such a correction by some simple experiments. However, what I would like to know is if there exist any software that can calculate the needed corrections. Is it possible by theoretical means to evaluate this quantitatively?
Yours sincerely
Paul Baggethun Manchester Materials Science Centre UMIST
} [snip] } } } Under some illumination conditions there may be no more than 1 beam electron } } } in the column at a time! } } With our HVEM (1.2 MV) and its ~3m column this turns out to be cor- } } rect--especially since we use low-dose conditions for ED work. } Wow! The current of electrons hitting the specimen must be quite low. } How do you estimate it? } Dear Philip, For the 100 micrometer C2 aperture, we get ~10^-11 amps, for the 30 micrometer C2 aperture (which is better because the spot at crossover is ~350 nm--the size of the P1 aperture) the current is too low to show up on the screen current meter or the second Faraday cage. We made a high-precision Faraday cage which goes on the bottom of the column, and we measure the beam current with an electrometer. Yours, Bill Tivol
About Macintosh-based software for image capture and manipulation -- We use the Prism image analysis & measurement package (Analytical Vision, Raleigh NC) together with a RasterOps graphics display & video image capture board on our Macintosh Quadra. We acquire color images with Adobe Photoshop. Works well and yes we're happy with it. (Usual disclaimer...)
Thor Bostrom
At 11:19 AM 1/17/96 -0500, Ruth Hughes wrote: } Hello, } I was wondering if anyone is using a Macintosh-based software for } image aquisition and manipulation that they are happy with? I have been } using Metamorph (Love it!) but now need something for the Mac. } Thanks in advance. } } Ruth Hughes } Central Institute for the Deaf } 818 S. Euclid } St. Louis, MO 63110 } e-mail RMH-at-CIDMV1.WUSTL.EDU } }
=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= Dr Thor Bostrom Analytical EM Facility Queensland University of Technology (QUT) GPO Box 2434, Brisbane, QLD 4001, Australia Ph: +61 7 3864-2351 FAX: +61 7 3864-5100 http://www.sci.qut.edu.au/aemf/ =-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
Nancy Desmond asked about signal averagers similar to the Dage DSP-2000. Our company has been making for many years a box called the OMNEX which is a stand-alone real time processor in some ways similar to the Dage.
The OMNEX has a lot of features. It can average, and integrate video frames. Integration comes in 4 flavors including some automatic modes. A unique feature is that it can control integrating cameras for long exposure in cases of low light. It can do background subtraction, dynamic subtraction, psuedocolor. It has a measurement section for linear measurements, area, path, counting, stopwatch. It has a built-in clock calender which may be displayed on the screen. It can zoom, too. 4 labels may be put on the screen. The whole thing is mouse controlled and extremely easy to use. There is even an onscreen help facility.
Very useful: complete digital contrast enhancement including histogram equalization and 4 custom lookup tables.
A very powerful feature is that it can do matrix operations like sharpening, gradient, edge detection, etc. Two color overlay is also possible.
It has a section to store 4 images and sequence them like a slide show (but up to 30 times per second.)
I am sure that there are other features that I am forgetting.
The Dage unit does not do most of these things. It is designed more for precise control of the video parameters like black level, etc.
The OMNEX costs $5350. It is also sold by Zeiss.
I can send you a brochure if you send along your address. Please call or Email with any questions.
We also have a very powerful video marking and measuring system called the XR-2000.
When I took my legally required training for radiation protection a few decades ago, I learned the (approximate) formula
d.r. = (0.3*(U**2)*I*Z)/r**2
where:
d.r. dose rate in [r/h] (roentgen per hour) U accelerating voltage in [kV] I electron beam current in [mA] Z (mean) atomic number of the target, r distance from the target in [cm]
Since practically all electrons that come out of the source are accelerated to U and hit something somewhere inside the instrument, setting I equal to the emission current (for which there is a meter on most instruments) will give you an upper limit on the electrons that produce x-rays. Most guns are designed for emission in the 0.1 A range. What most electrons hit is made out of stainless steel or brass, although some also hit Pt or W apertures, so setting Z=40 or 50 should be high enough to overestimate x-ray production by a bunch. Assuming that it takes, say, 8mm of steel to keep a small electron beam instrument from imploding under its own vacuum, we can calculate that under the worst case conditions no measurable x-rays can escape for "low voltage" work typical of SEMs. Although there are a lot of x-rays generated, the vacuum enclosure is necessarily a very efficient shield.
The instrument I work with, an ARL SEMQ microprobe, actually has a half-inch steel enclosure, and its 30kV power supply cannot be cranked up high enough to measure x-rays outside the instrument. Since we, too, have a law requiring annual inspections, we have verified by actual measurements under different conditions that this is true. To be on the safe side, the manufacturer has also specified that in the case of modifications no enclosure parts are to be made from aluminum (shielding efficiency goes with a high power of Z).
However, the above formula also tells you that the situation quickly changes if you use higher voltages. X-ray generation goes with the square of U, and I typically increases as well when the voltage increases, so that the "net" dose rate increases with something around 2.5 to 3rd power of U, and the shielding efficiency of metal decreases as the generated x-rays get more energetic. With 100 or 200 keV instruments, i.e. most TEMs, I would definitely start to worry about x-ray leakage, and require people to wear film badges or (perhaps preferably) dosimeter rings.
P.S.: Mid-Iowa temperature minus 9, wind chill index minus 60.
Alfred Kracher akracher-at-iastate.edu http://www.public.iastate.edu/~akracher
Message : I'm interested in suggestions for embedding media for polymeric thin films, typically automotive coatings, which will be cryomicrotomed in the -60 to -90C range. Most coatings will originate from a water based system.
I'm also interested in suggestions for cryomicrotomy courses which would include bulk and embedded polymeric materials (elastomers, coatings, alloys, and composites).
Thanks for your help.
Question :
How thin do you want the sections to be?
Paul Webster Center for Cell imaging Yale School of Medicine 333 Cedar Street New Haven, CT 06520.
I am interested in using image analysis to count bacteria, measure fungal hyphal lengths, and estimate bacterial and fungal biovolumes in soil samples. I am currently trying to use NIH-image for this purpose. If you are using NIH-image or another software package for this or a similar purpose, I would be interested in hearing from you. I am just getting started and have a number of questions.
Thanks,
Serita Frey
------------------------------------------------------------------------ Serita Frey Natural Resource Ecology Laboratory Colorado State University Ft. Collins, CO 80523 ------------------------------------------------------------------------
} This may be a naive suggestion but, wouldn't amorphous C work as well?
Dear John, It's not a naive suggestion; however, since I am applying the method for a 1.2 MV scope, the contrast is too low with a C film. I'm prepared for the possibility that even a Ge film might not be quite good enough. What I will need to do is to take an optical or computer trans- form of the image and find the zeros of the CTF, so the better the con- trast, the more accurate the results. Yours, Bill Tivol
As part of our action-oriented programs under our Affirmative Action Plan, this letter serves as notification that we are seeking minority and female candidates for the following open position, but all qualified candidates will be considered:
Senior Engineer, Mechanical Properties
- This position will be located at our Corporate Technology Center in Latrobe, PA, working in Research and Development.
- The Sr. Engineer position requires a Masters degree, with a Ph.D. preferred, in engineering science or materials related discipline and 2-8 years experience, dependent on the applicants degree, in the study of structure/property relationships. Effective communication, interactive and supervisory skills are required to work with other research personnel and the academic community on both short and long term R&D projects.
- This responsibilities for this position are:
Utilizing TEM (STEM), SEM, EDS, and related instrumentation, provides microstructural, physical property and mechanical property expertise on materials including but not limited to powder and sintered, carbide, cermet, and superhard materials.
Conducts mechanical property testing including tensile, compressive, fracture toughness, transverse rupture and wear at room and elevated temperatures.
Liaisons with outside organizations, universities and other laboratories involved in joint material related research programs.
Investigates and recommends purchase of new analytical instrumentation.
Conducts validation studies of testing procedures using SPC techniques and recommend solutions to resolve discrepancies.
Documents new and existing testing methods with specific procedures.
Represents Kennametal through technical presentations/publications, organizational memberships, serving on committees and elected offices.
- Kennametal Inc. offers an excellent relocation and benefits package and a salary range for this position from $3,920 per month to $5,880 per month dependent on the candidates qualifications and experience.
All candidates for whom resumes are submitted must acknowledge their referral source. Resumes should be sent directly to: Kennametal Inc. Attn: Dennis B. Harr Manager, Human Resource Services P.O. Box 231 Latrobe, PA 15650-0231 (412) 539-5434
Please respond ONLY to the above. DO NOT respond to the originator of this e-mail message.
Please ask your questions. I would be pleased to try to answer them as would others on the listserve.
John Libert
At 02:20 PM 1/19/96 -0700, Serita Frey wrote: } } I am interested in using image analysis to count bacteria, measure } fungal hyphal lengths, and estimate bacterial and fungal biovolumes in } soil samples. I am currently trying to use NIH-image for this purpose. } If you are using NIH-image or another software package for this or a } similar purpose, I would be interested in hearing from you. I am just } getting started and have a number of questions. } } Thanks, } } Serita Frey } } ------------------------------------------------------------------------ } Serita Frey } Natural Resource Ecology Laboratory } Colorado State University } Ft. Collins, CO 80523 } ------------------------------------------------------------------------ } }
} Hello ImageTool Users, } } First we would like to thank all of you for your patience and suggestions } for improving ImageTool. The reception for a image analysis and } processing package for Microsoft Windows has been great. We have } distributed over 1,000 copies world wide to major research institution } since our initial release on August 24, 1995. } } Extra! Extra! ImageTool NEW RELEASE Verion 1.1 } } Version 1.1 of ImageTool is presently available from our ftp site } ftp://maxrad6.uthscsa.edu The new version includes all bugs reported as } of 12/22/95 and some really nice features. We have developed a complete } object analysis and classification plug-in. This plug-in provides for } automated object counting and analysis with over 20 unique measurements. } These include: area, perimeter, x, y center, feret diameter, roundness, } compactness, major/minor axis length, slope and endpoints. Also included } are integrated density, binary and gray centroid. Using Objects } Classification, objects can be categorized into sub groups based on any } of the above object measurements. } } Also new in version 1.1 is support for the Data Translation DT3155 } High-Accuracy Monochrome PCI Bus Frame Grabber. The driver that is } shipping presently supports Windows NT and not Windows 95. Data } Translation will release the Win95 driver for this acquisition board in } February 1996. } } We hope that this new version will help many of you do your research. } Please keep those suggestions of new features and any bug reports coming. } Also please send information about how you are using ImageTool in your } research and what features you would like to see in the next release. We } hope to have a new release sometime in February. } } Thank you again for your support. } } S. Brent Dove } Diagnostic Sciences } University of Texas } Health Science Center } San Antonio, TX USA } Voice: (210) 567-3333 } Fax: (210) 567-3334 } Email: dove-at-uthscsa.edu } Web: ddsdx.uthscsa.edu } ftp: maxrad6.uthscsa.edu } } } ---------- } From: nih-image } To: Multiple recipients of list } Subject: Windows version } Date: Saturday, January 20, 1996 2:36AM } } Does anyone know if there is a Windows version of Image, or a program } similar. } } Thanks } } Candy } } }
Due to the government furlough, the deadline for submitting applications to NIST for postdoctoral positions has been revised as shown below.
POSTDOCTORAL POSITIONS AT NIST
The National Institute of Standards and Technology (NIST) is interested in receiving applicants for National Research Council (NRC) postdoctoral positions. In particular, applicants with interests in electron microscopy are welcomed for work in the Chemical Science and Technology Laboratory. Projects can be proposed in the fields of material science, chemistry, mineralogy or other related fields using a variety of techniques including high-resolution TEM, EDS, PEELS, holography, and electron energy loss imaging, etc.
INSTRUMENTATION AVAILABLE: A Philips CM300 FEG equipped with a Gatan Imaging Filter and a Philips CM30 with a PEELS unit are the primary TEM instruments. There are a variety of associated instruments available including scanning electron microscopes, secondary ion microprobes, x-ray diffraction units, etc.
SALARY: $45,500 per year (2 year appointments)
APPLICATION (WITH DRAFT OF PROPOSAL) DUE BY: February 1, 1996
SUPPORTING DOCUMENTS AND FINAL PROPOSAL DUE BY: February 15, 1996
EXPECTED STARTING DATE: Fall 1996 (PhD must be complete by this time)
FOR MORE INFORMATION: contact Eric Steel (301) 975-3902, eric.steel-at-nist.gov or Shirley Turner (301) 975-3923, shirley.turner-at-nist.gov.
I am looking for information on a company which I believe is called AMT. I have been told they make interface flanges for cameras on TEMs. In particular, I need to connect a camera to a 100CXII - either below the camera chamber or on the 35 mm port, and to a CM20 below the camera chamber.
The cameras will not be used for high resolution work, but for training purposes.
If anybody knows of the company or any other sources please let me know.
Keith Moulding.
Materials Characteristion Preparation Centre, Hong Kong Univeristy of Science and Technology,
In reply to Paul Webster from Yale about any Cryomicrotomy Courses. Suggest contacting David Remsen on e-Mail address dremsen-at-aaem.amc.anl.gov who is organizing a course on Cryomicroscopy and related techniques at Woods Hole MBL May 29-June5 1996.
Patrick Echlion Multi-Imaging Centre Cambridge University
On 19 Jan 1996, Paul Webster wrote:
} Message : } I'm interested in suggestions for embedding media for polymeric thin films, } typically automotive coatings, which will be cryomicrotomed in the -60 to -90C } range. Most coatings will originate from a water based system. } } I'm also interested in suggestions for cryomicrotomy courses which } would include bulk and embedded polymeric materials (elastomers, coatings, } alloys, and composites). } } Thanks for your help. } } Question : } } How thin do you want the sections to be? } } Paul Webster } Center for Cell imaging } Yale School of Medicine } 333 Cedar Street } New Haven, CT 06520. } }
Hi all, I am considering buying Image-Pro plus software to do some of the things a lot of you have mentioned in the past : feature counting on prints/images, digitalization of negatives, image enhancement and printing.
The system we are thinking of will be attached to a Nomarski microscope for optical work ( defect counting of etched wafer surfaces ), and to a seperate camera mounted on a light-box to digitalize negatives and prints. We have not made a final decision on : 1. resolution of cameras to buy. 2. black and white vs. color. 3. type of printer/s to have available ( the company suggests a Kodak for high quality work )
Anyone out there using a similar setup to count/size and digitalize ? Anyone using Image Pro Plus ?
thanks
Lucio
*********************************************************************** Lucio Mule'Stagno Physics Dept & Center for molecular electronics Univ. of Missouri- St.Louis tel 314 - 516 5933 fax : 314-516 6152 e- mail LUCIOM-at-NEWTON.UMSL.EDU
Hi all, I am considering buying Image-Pro plus software to do some of the things a lot of you have mentioned in the past : feature counting on prints/images, digitalization of negatives, image enhancement and printing.
The system we are thinking of will be attached to a Nomarski microscope for optical work ( defect counting of etched wafer surfaces ), and to a seperate camera mounted on a light-box to digitalize negatives and prints. We have not made a final decision on : 1. resolution of cameras to buy. 2. black and white vs. color. 3. type of printer/s to have available ( the company suggests a Kodak for high quality work )
Anyone out there using a similar setup to count/size and digitalize ? Anyone using Image Pro Plus ?
thanks
Lucio
*********************************************************************** Lucio Mule'Stagno Physics Dept & Center for molecular electronics Univ. of Missouri- St.Louis tel 314 - 516 5933 fax : 314-516 6152 e- mail LUCIOM-at-NEWTON.UMSL.EDU
It has been my experience that a camera mounted below the camera chamber subjects the final image to a magnification of at least twenty (20)times that of a film.If this is not desired I suggest you install the camera above the camera chamber where the magnification is a more reasonable five (5) times. Kate Connolly
In reply to Paul Webster from Yale about any Cryomicrotomy Courses. Suggest contacting David Remsen on e-Mail address dremsen-at-aaem.amc.anl.gov who is organizing a course on Cryomicroscopy and related techniques at Woods Hole MBL May 29-June5 1996.
Patrick Echlion Multi-Imaging Centre Cambridge University
On 19 Jan 1996, Paul Webster wrote:
} Message : } I'm interested in suggestions for embedding media for polymeric thin films, } typically automotive coatings, which will be cryomicrotomed in the -60 to -90C } range. Most coatings will originate from a water based system. } } I'm also interested in suggestions for cryomicrotomy courses which } would include bulk and embedded polymeric materials (elastomers, coatings, } alloys, and composites). } } Thanks for your help. } } Question : } } How thin do you want the sections to be? } } Paul Webster } Center for Cell imaging } Yale School of Medicine } 333 Cedar Street } New Haven, CT 06520. } }
I am looking for information about EELS analyser makers, for implementation on a 300 KV TEM. I heard of Gatan analyser or image filtering system, but would like to know if there are any other makers providing this kind of apparatus.
Thank you in advance for providing any fax number or E-mail address,
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Luis Sole i Sabaris E-08028 BARCELONA
Regarding magnification: The Gatan camera mounted below our HF-2000 and our 4000EX instruments has a magnification factor over the film of 1.44 times the width of the displayed image in inches (since the chip is about 1 inch on a side). For an image displayed on a hard-copy output that is say 7 inches wide, this give a final image magnification of 10 times the microscope magnification.
May I also add to this that I was not the one requesting information about cryomicrotomy or courses. I was responding to an anonymous contributor from Dupont (ratyrg-at-ESVAX.DNET.DUPONT.COM) including the original message with my question. Since then we have had a short discussion on the subject away from the BBS.
Someone in our depeartment would like to know what type of Mac they are using, or would recommend to use, to run NIH-Image?? They are typically dealing with images that are 1kbx1kb 24 bit colour. They are contemplating buying a new Mac to run Image and would like input as to what people are using and how well they run Image before buying a new computer. Any and all comments would be appreciated.
Mark Elliott Research Associate UBC-Pulmonary Research Lab
Detection of features in Nomarski (DIC) images can be tricky. Although your eye will see them, thresholding may be difficult because of the "shadowing effect" that DIC has. Even selection by color can be difficult. I had poor results thresholding transmitted light DIC (biological); better results with reflected light DIC (ceramics).
We use Optimas, an alternative to Image-Pro for Windows based image processing (color and b/w). You should also do a serious evaluation of frame grabber boards, including how well they work with your I.A. system. Our first system has an Imaging Technology CFG board, which was pricey but worked well. Our second system has a Coreco TCX, which has some timing delays with Optimas.
No commercial endorsement should be implied.
David Rothbard
} Hi all, } I am considering buying Image-Pro plus software to do some of the } things a lot of you have mentioned in the past : feature counting on } prints/images, digitalization of negatives, image enhancement and printing. } } The system we are thinking of will be attached to a Nomarski } microscope for optical work ( defect counting of etched wafer surfaces ), } and to a seperate camera mounted on a light-box to digitalize negatives and } prints. We have not made a final decision on : } 1. resolution of cameras to buy. } 2. black and white vs. color. } 3. type of printer/s to have available ( the company suggests a Kodak } for high quality work ) } } Anyone out there using a similar setup to count/size and digitalize ? } Anyone using Image Pro Plus ? } } thanks } } Lucio }
I'm at present trying to visualize through TEM an atypical form of growth of a dimorphic fungus, Mucor rouxii, which under normal conditions grows as mycelia and when cAMP analogs are added to the growth media grows iso- diametrically; germ tube emission is impaired, but growth is not impaired. Cells , under these growth conditions are very fragile, and even tend to explode when being under a coverslip. The cell wall is several times wider than the normal cell wall. The microscopic preparations for TEM are not good; the cell looks completely collapsed and detached from the wall. We have already tried adding sorbitol 0.5 M and KCl 0.6 M at the end of the growth period, before isolating the cells, with no success. Can someone give me a suggestion, or indicate me a reference . We obviously are no experts in electronic microscopic observations. Thank you very much. -- Silvia Moreno smoreno-at-kinase.uba.ar
Does anyone in this list know the e-mail address or the FAX number of prof. Ernst H. K. Stelzer at European Molecular Biology Lab? Please reply to me directly. Thanks!
About a year ago, after using EM's for 27 years and having developed cataracts in both eyes at an unusually early age (49), I asked the question through this medium about eye damage resulting from long-term use of electron microscopes. As in the current discussion some interesting comments were made. There is definitely evidence that radiation from a variety of sources causes cataracts, typically in the posterior capsule of the lens - not the nucleus where senile cataracts usually develop. However, modern electron microscopes should be adequately screened to prevent this. In my case the cataracts were apparently typical of radiation-induced cataracts although when and where the irradiation took place is anyone's guess. Someone has mentioned UV radiation from vacuum evaporators. This could be a possibility because I know of little in the way of precautions which are routinely taken to prevent this.
Robin Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za (tel: +27 461 318168 - fax: +27 461 24377)
Subject: Time: 7:20 AM Charging for Services Date: 1/23/96
This request is made in the context of a U.S. university laboratory run as a 'recharge center' with a 'revolving account' where most of the users are funded by NSF or NIH. The laboratory provides a range of services in light and electron microscopy for researchers from several departments. Service contract costs for the instruments are recovered through hourly usage charges. Consumable supplies are charged at replacement cost or are replaced 'in kind' by the users. Other services are provided at cost. The university requires that, over time, the account will not make a profit and will not carry a continued balance greater than 10% of the operating costs of the facility.
There has been increasing interest in devising a system whereby users could be charged a fixed sum that would provide 'unlimited' access to one or more instruments/services for a specified time. My understanding is that such a system would violate granting agency prohibitions against prepayment for services, and would similarly violate current university regulations governing recharge centers.
I am interested, therefore, in hearing from anyone who has a system in place that provides in some way for 'lump sum' payments for instrument access/laboratory services. Since the university's recharge regulations will be revised this year, I would also like to hear from anyone whose laboratory appears to operate under less stringent university regulation.
Thank you.
Edward J. King king-at-bioscience.utah.edu Department of Biology University of Utah
We are looking for algorithms to process EDS spectra. The algorithms should include background subtration and calculation of k-ratios from unknown specimens (standardless analysis). Any information would be very helpfull.
Joe Geller Geller MicroAnalytical Lab. 426e Boston St. Topsfield, Ma 01983-1212 508 887-7000 fax 887-6671
The Electron Microscopy Center at Texas Tech University Health Sciences Center has an EM Technician II position open. Basic requirements are a bachelor's degree and one year experience with EM. The bachelor's degree can be substituted with an associate's degree from a 2 year EM program or a MSA accreditation and 3 years of experience. Any combination of experience in EDXA, morphometry, image analysis, light microscopy, confocal, histology, and/or flow cytometry would be a plus. Please reply before January 31, 1996 to: TTUHSC Office of Human Resources, 3601 4th Street, Lubbock, Texas 79430 TTUHSC IS AN EEO/AA EMPLOYER
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
The PowerMacs (PPC601 or 604 based) are generically the best way to go for running NIH Image. I have been using the 7500 (which is a PCI-bus based system) and the PCI Scion LG-3 frame grabber (PAL version). The PCI version is very fast for video capture, although it sounds like your user has an alternate source of images. (Also, they need to be aware that NIH- Image is not a true 24-bit color package - It handles the three color planes as a stack of 8-bit images. They should consider something like IP- lab Spectrum for true-color quantification or Photoshop for "graphic arts type" image massaging and printing.) I have also used the built-in video on the 7500. It is a reasonable image quality, but lacks the "scientific" capabilities that the Scion has (grayscale quality, gain/level adjust, analog out, digital I/O, etc.).
The 7500/8500/9500 PowerMacs all have high-speed internal SCSI, so disk I/O is very good. The 7500 is probably the best "bang/buck" system right now. It also uses a daughterboard processor card, so it can be upgraded to a PPC 604, in principal. If I had the budget and had to spend it right now, I would look very hard at the 132 MHz 9500. Of course, this family has now been out long enough that it is likely Apple will introduce a new even- faster Mac family soon. My impression has been that the best value has been on the model which is two steps back and within the same family as the most-recent top model (Current model example: high end is the 9500, hence the 7500 is the best value - i.e., affordable on tight budget, but excellent performance. Most recent past example: the 950 high-end/650 best buy). The clones have some VERY IMPRESSIVE specifications, (combination PCI/NuBus, high processor speed, etc) but may require optional hardware that is already in the Mac, depending on your use environment.
Hope this helps
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
As I am in one of the facilities who used to have a "pre-paid, lump-sum" system in place that appeared to make everybody happy, only to have a federal audit come in and SEVERLY penalize us for doing so, I would also be interested in what other recharge facilities are doing. Please keep this dialog going, or post a compilation of replies!
Mahalo, Tina
***************************************** Tina (Weatherby) Carvalho * Biological Electron Microscope Facility * University of Hawaii * (808) 956-6251 * tina-at-ahi.pbrc.hawaii.edu * http://www.pbrc.hawaii.edu/bemf/ * *****************************************
Years ago John Russ published a couple of dozen algorithms for carrying out various tasks involved in processing EDS spectra in the "EDAX EDITor", a publication of the EDAX Company. Although these are now several years old, they still might be useful as a starting point, and are certainly of some historical interest. If you are interested in them, John can probably help you with this; otherwise, I still have copies of a number of issues of the EDAX EDITor, and could let you borrow them. W. C. Bigelow (bigelow-at-umich.edu)
Look at the nih-image web site, http://rsb.info.nih.gov/nih-image/ which you can also find using "Net Search" for "nih image", in your favorite web browser like Netscape.
This is an interesting web site even if you don't want to use NIH Image.
(NIH Image runs on power macs as well as the 68k macs).
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
Someone wrote recently:
"I'm interested in suggestions for embedding media for polymeric thin films, typically automotive coatings, which will be cryomicrotomed in the -60 to -90C range. Most coatings will originate from a water based system"
Could you be more specific as to whether you are interested in the dispersion of additives within the thin film (of which there are typically high loadings in many "automotive" coatings) or if you are interested in the air interface surface detail. In either case, you do have to think about "passivating" the top (and bottom) surfaces in order to keep the embedding resin, what ever it is you eventually use, from possibly swelling or other wise changing in some subtle way the polymer layer of interest. We have found that a thin film of sputtered gold works fine, Pt slightly better. For the opposite size, we like to coat with Al so that there is never any question as to which side is which once in the TEM.
If interested primarily in the dispersion of additives in the layer, then the best approach is to embed completely the now both-side passivated film and section.
If interested primarily in the top layer, we "back embed" only (e.g. the bottom surface but after Al coating), gold sputter the top surface but do not embed the top surface. That way, by following the gold layer one can fairly easily discriminate between fact and artifact, that is, surface disruption that was there to begin with vs. surface disruption caused by the knife. We find that additive particles tend to be more likely to be pulled out of the section when using this variation of the approach.
We have found overall several of the so-called "Epon substitutes" are ideal for this particular application, such as our own "SPI-Pon 812" resin. Since the hardness of the final block has to be made to reflect the "hardness" of inorganic additives, if present, the "Epon 812" type systems permit what we find to be the very widest latitude in terms of hardness levels. The fact the system was once "water based" should not make any difference in terms of your choice of embedding system since by the time the resin "sees" the sample, any water has long since disappeared. If your sections do not come out perfect that first time, then varying the hardness of the block might be the first next thing you would want to try. But don't lose confidence in the resin system.
And assuming you are the cost conscious type, the "Epon 812 substitute" resins are about the lowest cost of any resin.
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: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
We have the following 23 filaments which were purchased for an ETEC Autoscan, they are free to anyone who would like them for an ETEC or similar instrument: "EBTEC rebuilt filaments for ETEC, MR 73" (Probing and Structure) - 13 filaments "VL-EO-R" (Energy Beam Sciences) - 10 filaments
Regards
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
Message-Id: {199601232339.AA19555-at-lucy.swin.edu.au} Comments: Authenticated sender is {hbrinkies-at-gpo.swin.edu.au}
This message might be mainly of interest to the Australian EM community, however, I did not receive any reply to my advertisement in the latest Australian EM Newsletter (48).
So let's try again:
For sale (very cheap). Preloved ED-System. Ortec EEDS-II (needs attention). Consisting of Si(Li) horizontal detector (refurbished by HNU Systems in 1993). Two consoles: Mark I (1981) + Mark II (1983). Epson Printer. Software on 8" floppies, circuit diagrams, manuals, spare 8" disk drives, several spare boards. Best offer accepted.
Hans G Brinkies SWINBURNE, University of Technology Mechnical and Manufacturing Engineering P.O.Box 218 - HAWTHORN, 3122 - Australia Phone: + 61 3 9214 8657 Fax: + 61 3 9214 8264
The Electron Microscopy Center at Texas Tech University Health Sciences Center has an EM Technician II position open. Basic requirements are a bachelor's degree and one years experience with EM. The bachelor's degree can be substituted with an associate's degree from a 2 year EM program or a MSA accreditation and 3 years of experience. Any combination of experience in EDXA, morphometry, image analysis, light microscopy, histology, and/or flow cytometry would be a plus. Please reply before January 31, 1996 to: TTUHSC Office of Human Resources, 3601 4th Street, Lubbock, Texas 79430 TTUHSC IS AN EEO/AA EMPLOYER
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
If not, please update your e-mail address list with my new location:
dbd1-at-uclink4.berkeley.edu
Thank y'all.
Doug Davis EML Berkeley
=F8=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5= =B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=F8 =F8 Doug Davis =F8 =F8 Staff Research Associate =F8 =F8 Electron Microscope Facility =F8 =F8 University of California =F8 =F8 Berkeley, CA 94720 =F8 =F8 (510) 642-2085 =F8 =F8 dbd1-at-uclink4.berkeley.edu =F8 =F8=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=B5= =B5=B5=B5=B5=B5=B5=B5=B5=B5=B5=F8
To do this experiment correctly you must energy filter the diffraction pattern to correctly determine the diffracted intensities. This filtering removes all the inelastic scattering. It is not sufficient to simply measure the intensity just inside the ring to correct for "background". You had best check the literature before doing alot more work.
There are some very good papers by David Cockayne & Colleagues at the University of Sydney. Circa late 1980's in Acta Crys.
I would suggest you look them up before continuing.
Message-ID: {MAPI.Id.0016.00683536202020204434454530303237-at-MAPI.to.RFC822} Read-Receipt-To: Jean-Louis Beek {ah56-at-solo.pipex.co.za} Priority: Normal To: microscopy-at-Sparc5.Microscopy.Com MIME-Version: 1.0
The Association Vaudoise des Chercheurs en Physique organizes a
**** Winter School****
M=E9thodes modernes de diffusion et de diffraction des neutrons, =E9lectrons= et RX
in Grimentz (Valais) Switzerland (nice for skiing, isn't it?) Feb. 25 to March 2 1996
It is intended for people who are not specialized in (all) these methods and who would like to get a general knowledge of the similarities, the differences and the potential applications of these techniques. You can get all the information on Internet
http://cimewww.epfl.ch/avcp/index.html
The lectures will be approx. 60% in French and 40% in English (25 hours at total, 15 lecturers) The deadline for registration is Feb. 3
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch // pabuffat-at-cimesg1.epfl.ch ______________________________ Eudora F2.1 ___________________________
The Association Vaudoise des Chercheurs en Physique organizes a
**** Winter School****
M=E9thodes modernes de diffusion et de diffraction des neutrons, =E9lectrons= et RX
in Grimentz (Valais) Switzerland (nice for skiing, isn't it?) Feb. 25 to March 2 1996
It is intended for people who are not specialized in (all) these methods and who would like to get a general knowledge of the similarities, the differences and the potential applications of these techniques. You can get all the information on Internet
http://cimewww.epfl.ch/avcp/index.html
The lectures will be approx. 60% in French and 40% in English (25 hours at total, 15 lecturers) The deadline for registration is Feb. 3
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch // pabuffat-at-cimesg1.epfl.ch ______________________________ Eudora F2.1 ___________________________
I am looking for the fax number (or even E-Mail address) of the company AREMCO products, OSSINING,NY. They sell Aremco crystalbond 509 glue, that we use for preparing samples.
thank you in advance
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Luis Sole i Sabaris E-08028 BARCELONA
Please complete/return form below to remain/get on our lists.
Scanning Microscopy International Post Office Box 66507, Chicago (A.M.F. O'Hare), IL 60666-0507, U.S.A. Telephone: (708) 529-6677 / FAX: (708) 980-6677 E.mail: 73211.647-at-compuserve.com
Scanning Microscopy 1996 meeting May 11-16, 1996, Bethesda, Maryland (suburb of Washington, DC)
Symposium on: Scanning Probe Microscopies and Related Techniques for the Biological and Materials Sciences
Note: A number of SPM papers will also be presented durin the program on "Pattern Formation and Nanoscaled Structures in Thin Film Formation" at the same time at same venue. THOSE PAPERS ARE NOT LISTED BELOW. Please request / see a separate flier on that.
Program organizers: Dr. David P. Allison, Oak Ridge Natl. Lab., TN (E-mail: allisondp-at-ornl.gov); Prof. Chunli Bai, Chinese Acad. Sci., Beijing (E.mail: clbai-at-infoc3.icas.ac.cn); Prof. Lawrence A. Bottomley, Georgia Inst. Tech., Atlanta (E.mail: lawrence.bottomley-at-chemistry.gatech.edu); Prof. Masamichi Fujihira, Univ. Yokohama, Japan (phone: 81 22 2152021 / FAX: 81 22 2152020 / E.mail: mfujihira-at-bio.titech.ac.jp); Dr. Heinrich J.K. Hoerber, European Molec. Biol. Lab., Heidelberg, Germany (E.mail: hoerber-at-embl-heidelberg.de); and Prof. Douglas J. Thomson, Univ. Manitoba, Winnipeg, Canada (E.mail: thomson-at-ee.umanitoba.ca).
Following up on the past SPM meetings, this symposium will provide a nice occasion to present novel discoveries as well as reviews of recent developments in theory, instrumentation, and applications of scanning tunneling microscopy and related techniques, including atomic force microscopy, magnetic force microscopy, near field optical microscopy, etc. Applications of STM and other scanning probe techniques should emphasize the studies of adsorbates as well as physical and chemical process at solid surfaces. Topics of interest include: studies of processes on metal, semiconductor, and other solid surfaces: imaging of molecules, especially biomolecules; imaging of cells and other biological structures; tip-induced effects; etc.
Papers can still be offered: please contact one of the organizers or Dr. Om Johari at Scanning Microscopy International.
List of presentations (as of January 23, 1996) in alphabetical order
M.J. Allen, Digital Instruments, Santa Barbara, CA et al.: The Chromatin Structure of Well-Spread Demembranated Human Sperm Nuclei Revealed by AFM
D.P. Allison et al., Oak Ridge National Lab., TN: High Resolution Physical Mapping of EcoRI restriction Sites on Intact Cosmids by AFM Imaging
P.C. Zhang, C. BAI, P.K.H. Ho, Q. Li, Y. Dai, Y.S. Wu, B.F. Sheng, Chinese Acad. Sci., Beijing: AFM Study of Interactions Between Tumor Necrosis Factor and Its Monoclonal Antibodies
J. Bereiter-Hahn et al., Univ. Frankfurt, Germany: Regulation of Cell Surface Motility, as Revealed by Scanning Acoustic Microscopy
G. Collins, Topomatrix, Santa Clara, CA: (1) A Novel High Resolution NSOM; (2) Polymer Science Applications of a Scanning Thermal Microscope Having a Resistively Heatable Probe; (3) Applications of a Combined SPM/SEM
E.D. Dahlberg et al., Univ. Minn, Minneapolis: Review: Magnetic Force Microscopy and Magnetotactic Bacteria
O. Enea, Univ. Poiters, France: Topographic Studies of TiO2 and SnO2 Ceramics by Environmental SEM, SEM and AFM
R. ESCHRICH, Max-Planck-Inst. Biochem., Martinsried, Germany; G.L. Kumar, T.A. Keil, R. Guckenberger: AFM on the Olfactory Dendrites of Giant Silkmoth Antheraea
M. Fujihara, Tokyo Inst. Tech., Japan: Review: AFM of Solid Surfaces in Aqueous Solutions
D. Goddard, Br. Nucl Fuels, Preston, UK: AFM of Bacterial Biofilms with Application to Microbially Influenced Corrosion
H. Hansma, Univ. Calif., Santa Barbara: Review: Atomic Force Microscopy of Biomaterials
M. HARA, W. Knoll, RIKEN, Saitama, Japan: Review: STM and AFM Studies of Self-Assembled Monolayer Growth
D.O. HENDERSON, Y.S. Tung, R. Mu, Fisk Univ., Nashville, TN; W.A. Curby, M. Mercado, Fed. Aviation Rech. Ctr., Atlantic City, NJ: Optical and Atomic Force Microscopy of Pentaerythritol Tetranitrate Nanoclusters on Si(100)
E. Henderson, Iowa State Univ., Ames: Biomolecular Detection with the AFM (Tentative Title)
H. Hoerber, European Molecular Biology Lab., Heidelberg, Germany: Measuring Surface Forces with the AFM
D. HUANG, Y. Yamamoto, Yamamoto Quantum Fluctuation Project, Tokyo, Japan and Stanford Univ., CA: Hydrogen Atom Extraction and Redeposition on the Monohydride Si(100)-2x1:H Surface
A. Ikai, Tokyo Inst. Tech., Japan: Review: Measurements of Mechanical Parameters of Proteins and Chromosomes with AFM
M.D. JOHNSON, Univ. Oklahoma, Norman, and H.W.M. Salemink: Review: Cross-Sectional STM on Semiconductor Heterostructures (to be presented during the Semiconductors program)
G.L. KUMAR, Max-Planck-Inst. Verhaltenphysiol., Seewiesen, Germany; R. Eschrich, R. Guckenberger, T.A. Keil: In Search for Putative Pheromone Receptors on the Membrane of Olfactory Dendrites in Silkmoths (A. polyphemus and A. pernyi) Using the AFM and SEM
L. Kuutti, VTT Biotech. Food Res., Espoo, Finland: Identification and Surface Structure of Crystalline Cellulose Studied by AFM
G. LEE, A.D. MacKerell, L.A. Chrisey, R.J. Colton, US Naval Res. Lab., Washington, DC: Measuring Inter- and Intramolecular Forces in Biomolecules
Y. Lyubchenko, Arizona State Univ., Tempe: AFM Studies of RecA-DNA Complexes
J.F. Marchiando, NIST, Gaithersburg, MD:: Methods for Interpreting Measurements from a Scanning Capacitance Microscope (to be presented during the Semiconductors program)
M. McDermott, Univ. Alberta, Edmonton, Canada et al.: Review: Chemical Mapping with Force Microscopy
M. Rivera, M. MILES et al., Univ. Bristol, UK: Phase Transitions in Liquid Crystals Observed by SPM
W. MIZUTANI, M. Motomatsu, H. Ogiso, H. Tokumoto, Nat. Inst. Adv. Interdiscpl. Res., Tsukuba, Japan: STM with Local Non-Linearity Detection of Organic Thin Films and Ion Irradiated Graphite
D.J. MUELLER, F. Schabert, A. Engel, Univ. Basel, Switzerland: Review: Structural Changes of Native Membrane Proteins Monitored at Subnanometer Resolution with the AFM
H. MURAMATSU, Seiko Instruments, Chiba, Japan; T. Ataka, N. Chiba, K. Nakajima, M. Fujihira Review: Fluorescence Imaging and Spectroscopy of Biomaterials in Air and Liquid by SNOM/AFM
P. Nagy, MTA KFKI Res. Inst., Budapest, Hungary: Review: SPM Image Reconstruction
R. Perez, Univ. Cambridge, UK: First Principles Simulations of Atomic Resolution in Non-Contact AFM (to be presented during the Fundamental Physics Program)
R. RAITERI, H.-J. Butt, Max-Planck-Inst. Biophy., Frankfurt, Germany: Changes in Surface Stress Measured with an AFM
B. Samori, Univ. Calabria, Bologna, Italy: DNA Imaging by SFM (exact title to come)
W. Haiss and J.K. SASS, Fritz-Haber-Inst., Berlin, Germany: STM Surface Stress Measurements in Electrochemistry
Y. Shirane, Univ. Tokushima, Japan: Surface Observation of Calcium Oxalate Monohydrate Crystals by FE- SEM and AFM (to be presented during the Stones and Crystals program)
R.P. Singh, Indian Min. Sci. Tech., New Delhi: STM and Application Possibilities
V. Snitka et al., Kaunas Univ., Lithuania: Acoustic Waves Investigation by Atomic Force Microscopy
I. Stangel, McGill Univ., Montreal, Canada: The Applications and Limitations of AFM to Complex Biomaterials Surfaces: Effects of Acid Demineralization on Human Dentin
M.S. UNLU, B.B. Goldberg, Boston Univ., MA: Review: Characterization of Materials and Devices by Near Field Optical Scanning Microscopy (to be presented during the Semiconductors program)
J. Vesenka, Calif. State Univ., Fresno: Review: The Diameter of Duplex and Quadruplex DNA Measured by SPM
VU THIEN BINH, F. Feschet, V. Semet, S.T. Purcell, Univ. Lyon, France: Fresnel Projection Microscopy: Theory and Experiment: Electron Microscopy with Nanometer Resolution at ~ 200 eV
J.W. Wu, J.H. GU, J.H. Fang, Z.H. Lu, Southeast Univ., Nanjing, China: A New Phenomenon at a Small Gap Resistance When Using STM to Modify Gold Surface
T. WU, Z. Ai, Southeast Univ., Nanjing, China: Autostereogram: A New Method for Scanning Probe Microscopy
Z.D. Xiao et al., Southeast Univ., Nanjing, China: A New STM System Using Radioactive Tip
F. Zenhausern, IBM Watson Research Ctr., Yorktown Hts., NY: New Developments in Near Field Optical Microscopy and Spectroscopy
---
Other related programs at the same venue (*: Fliers available):
*Fundamental Physics in Microscopy and Microanalysis,
*Pattern Formation and Nanoscaled Structures in Thin Film Formation
*Scanning Microscopy and Semiconductors: Metrology and Diagnostics;
Several Biological programs including: Microanalysis and Imaging, *Immunolabelling, *Radiation Effects, Apoptosis, *Dentistry, Corrosion Casting, *Inner Ear, *Bone Biology, *Stones and Crystals, etc.);
Several Biomaterials Related Programs organized under "*Cells and Materials": (1) Skeletal Tissue / Biomaterials; (2) Foreign Body Reactions; (3) Biointerfacial Reactions at Biomaterial Surfaces; (4) Innovative Drug Delivery Systems; (5) Blood-Related Biomaterials; (6) and (7) Dental Biomaterials.
and Food Structure
----------------
For more information, please complete and return the form below to
__ I wish to present at the Scanning Microscopy 1996 meeting (tentative title and summary on a separate sheet), please send a Letter of Intent form.
__ I cannot present, but am likely to attend the 1996 meeting, please keep me informed and send me: ___ 1996 Registration / Hotel form.
__ I can neither present nor attend; please add/keep my name on your mailing list. ___ Send me a mailing list form.
Please send: 1996 program fliers (*list programs here):
___ Instructions for Authors / ___ Major subject index / Table of Contents for: ___ Scanning Microscopy / ___ Cells and Materials / ___ Food Structure;
Flier on Scanning Microscopy:
___ Supplement 6, 1992 ("Signal and Image Processing in Microscopy and Microanalysis");
___ Supplement 7, 1993 ("Physics of Generation and Detection of Signals Used for Microcharacterization");
___ Supplement 8, 1994 ("Science of Biological Microanalysis")
___ Flier on the special issue: Interface Formation and Dynamics in Layered Structures (Scanning Microscopy, Vol. 8, no. 4, 1994).
___ Flier on 1996 Pfefferkorn Conference on Electron Image and Signal Processing, May 18-22, 1996 at Silver Bay, New York
Scanning Microscopy International P.O. Box 66507, Chicago (AMF O'Hare), IL 60666, USA Telephone: (708) 529-6677 / FAX: (708) 980-6677 E.mail:73211.647-at-compuserve.com
Fifteenth Pfefferkorn Conference on Electron Image and Signal Processing
May 18-22, 1996 at Silver Bay Association, Silver Bay, NY
The Conference will provide a forum for discussion of the current status, recent advances and prospects of many aspects of image and signal processing in electron and near-field microscopy and microanalysis. The principal themes are three-dimensional reconstruction, image restoration, enhancement and analysis (including morphological and algebraic approaches in particular), electron holography, the phase problem, processing image sets (from STEM detectors and from defocus and tilt series, for example), image simulation, and hardware and software for image acquisition and processing and for microscope control. Please see below for a list of speakers.
The organizers are: Dr. Peter W. Hawkes, CNRS Lab. Opt. Elect., B.P. 4347, 31055 Toulouse Cedex, France (Phone: 33-62-257884 / FAX 33-62-257999 / E.mail: hawkes-at-cict.fr); Dr. W. Owen Saxton, Univ. Cambridge, U.K. (FAX: 44-1223-334 567 / E.mail: wos1-at-cus.cam.ac.uk); and Dr. Joachim Frank, NYS Dept. Health, Wadsworth Center, Albany, NY 12201-0509, USA (Phone: 518 474 7002 / FAX: 518 474 8590 / E.mail: joachim-at-wadsworth.org). Interested contributors should contact either of the organizers with a title and a one page summary; please also include other relevant information (e.g., time desired, reprints of relevant publications, etc.).
Full-length papers will be published in the Conference Proceedings to be issued as Scanning Microscopy Supplement 11, 1997.
Conference Information: Registration begins at 6 PM on Friday, May 17. The first lecture will be at 8:30 AM on Saturday, May 18; the Conference will end at lunch on Wednesday, May 22. Sessions will be daily in mornings (from 8:30 to 1), afternoons (about 4:30-7), and evenings (about 8:45 to 10). Ample discussion time will be allowed; active participation by attendees is invited. Because of the limited number of attendees, the Pfefferkorn Conferences provide an excellent opportunity for in-depth discussions and personal contacts. Conference attendance is by application or invitation only. Qualified registrants will be accepted on a first-come-first-served basis. To apply: please submit name, mailing and E.mail addresses, phone (work and home) and FAX numbers, and a 50-100 statement about your qualification relative to the theme (and topics) of the conference, accompanied by the full registration fee of US $200 (includes attendance to sessions, coffee-breaks, and a copy of the Conference Proceedings).
Conference Location: The sessions will take place at one of the conference rooms at Silver Bay Association (SBA), Silver Bay, NY 12874 (phone: 518 543 8833 / FAX: 518 543 6733 / E.mail: sbaconf-at-aol.com), located 80 miles north of Albany, NY. This 600 acres beautiful, peaceful, registered historical place, located on Lake George and in the Adirondack mountains, offers dozens of water and land sports, gym programs, hiking, etc. and is filled with gardens, rolling lawns and gentle streams. It is ideal for a family vacation or retreat. The room rates (all rates include three full meals daily!) range from $40 per person (in two and four bed-room cottages ideal for four and eight persons, respectively), to $55 and $65 per person (based on two persons to a room) in rooms without and with private baths, respectively. Accompanying children aged 4 to 12 pay half of these rates. Single rooms are available at a cost of $69 (without private bath) or $80 (with private bath). It is expected that all attendees will stay at this location. To make accommodation reservations, please contact SBA directly and inform us of your travel details (mode and day/time). Details about getting to Silver Bay will be sent to all attendees: nearest airports are Albany (New York) and Burlington (Vermont); foreigners may find it easier to fly to Montreal, Canada (about 150 miles north) or New York (about 200 miles south). There is one daily train each way from New York and Montreal which stops in Ticonderoga, NY (about 20 miles from Silver Bay); some bus service is available too. Appropriate van transportation from Albany (at costs ranging from $50 per van for less than 4 people, or $10 per person for more than 5 persons) will also be arranged by SBA. Attemps will also be made to form car pools with those driving.
Past Conferences: Started in 1982, in honor of the late Prof. Gerhard E. Pfefferkorn, these Conferences present an in-depth discussion of fundamental topics in scanning microscopy and related techniques. Two conferences on topics directly related to the theme of the 1996 Conference were held in 1987 and 1991; registrants to the 1996 Conference can obtain those proceedings [containing original peer-reviewed, full-length papers published as Scanning Microscopy Supplements 2 (1998) and 6 (1992)] at special package prices (including uninsured cheapest rate mail delivery) of $95 (for US delivery) and $105 (for outside US delivery).
Scanning Microscopy International (SMI), a not-for-profit organization, sponsors the annual Scanning Microscopy, Cells and Materials, and Food Structure meetings: 1996 meeting will be from May 11 to 16 at Hyatt Regency Hotel, Bethesda, Maryland (suburb of Washington, D.C.]; publishes the international journals: Scanning Microscopy (previously Scanning Electron Microscopy till 1986), Food Structure, Cells and Materials (from 1991), and Scanning Microscopy Supplements (Proceedings of the Pfefferkorn Conferences); and issues publications devoted to selected topics derived from its journals. Details of the programs planned for the Scanning Microscopy 1996 meeting, a complete current list of publications, samples Tables of Contents of publications, etc. are available on request. The topics of this 1996 Conference are relevant to the themes of our journals; papers for publication are invited (Instructions for Authors are included in our journals or available on request).
For more information, please contact Dr. Om Johari, at SMI.
---
List of speakers as of January 22, 1996 (note: * = invited but not formally accepted as yet; when there are multiple authors, speakers name is in CAPITALS).
G. ADE, Physikal.-Techns. Bundes., Braunschweig, Germany: (1) A Digital Method for Noise Reduction in Holographic Reconstructions and Electron Microscopical Images (2) Coma-Free Alignment of Electron Microscopes and Digital Determination of Aberration Coefficients (with R. Lauer)
G. Anstis, Univ. Technol., Sydney, Australia; C.R. Birkeland, S.C. Anderson, D.J.H. Cockayne: Computation and Quantitative Analysis of HAADF Lattice Images of GaAs
N. Boisset, J.C. Taveau, V. You, F. de Haas, J. Lamy, URA CNRS, Tours, France: Refinement of Single Particle 3D Reconstruction (Art Method) Based on Topological Selection of EM Views
N. Bonnet, Univ. Reims, France: Multi-Dimensional Spectrum and Image Processing
C.B. Boothroyd, Univ. Cambridge, U.K.: Recent Results on Energy Filtered Image Quantification
C. BURMESTER, Max-Planck-Inst. Mol. Physiol., Dortmund, Germany; K.C. Holmes, R.R. Schroeder (Max-Planck-Inst. Med. Forschung, Heidelburg, Germany): Feasibility of the Multiple Isomorphous Replacement Method in Protein-Electron-Diffraction
H. CHENG, T. Baker, Purdue Univ., W. Lafayette, IN: The Correlation Approach to Virus Phasing
*M. Coster, Univ. Caen, France: Morphology and SEM Image Analysis: Recent Progress
C. DINGES, H. Rose, Tech. Hochsch., Darmstadt, Germany: Simulation of TEM Images and Diffraction Patterns Considering Phonon and Electronic Excitations
J. Frank, NYS Dept. Health, Albany, NY: Three-Dimensional Reconstruction (exact title to come)
S. Swaminathan, I.P. Jones (Univ. Birmingham, UK), N.J. Zaluzec (Argonne Natl. Lab.), D.M. Maher (No. Carolina State Univ., Raleigh), H.L. FRASER, Ohio State Univ., Columbus: Determination of Accurate Low Order Structure Factors for Si and TiAl Using Energy Filtered CBED
D.R. Beniac, G.J. Czarnota(1), F.P. Ottensmeyer(1), G. HARAUZ, Univ. Guelph, Canada; (1 = Ontario Cancer Institute, Toronto): Challenges of Three Dimensional Reconstruction of Nucleoprotein Complexes from Electron Spectroscopic Images
P. Kruit, B.M. Mertens, Delft Univ. Technol., Netherlands: Image Acquisition by Scanning in Fourier Space
M.K. KUNDMANN, Gatan EELS Software, Downers Grove, IL; S.L. Friedman, A.J. Gubbens, O.L. Krivanek: Characterization and Correction of TEM Energy Filter Aberrations
P.L. Bellon, S. LANZAVECCHIA, Univ. Milan, Italy: The Moving Window Shannon Reconstruction in Real and Fourier Domain. Applications in Tomography
H. Lichte, Univ. Dresden, Germany: Pitfalls on the Road to 0.1 nm with Electron Holography
M. MANKOS, IBM Watson Res. Ctr., Yorktown Hts., NY; M.R. Scheinfein, J.M. Cowley (Arizona State Univ.): Holography and the STEM
M.R. McCartney, Arizona State Univ., Tempe: Recent Applications of Electron Holography
D.G. MORGAN, D.J. DeRosier, Brandeis Univ., Waltham, MA: Analysis of Disordered Helices Using Correlation Methods
P.D. NELLIST, S.J. Pennycook, Oak Ridge Natl. Lab., TN: Probe and Object Function Reconstruction in Incoherent STEM Imaging
M.A. O'Keefe, Lawrence Berkeley Lab., Univ. Calif., Berkeley: On-Line Remote-Control Electron Microscopy with Emphasis on the Image and Signal Processing
P.A. PENCZEK, J. Zhu, R. Schroeder, J. Frank, NYS Dept. Health, Albany, NY: Three-Dimensional Reconstruction with CTF Compensation from Focus Series
T. Plamann, Univ. Bristol, U.K.: Three-Dimensional Propagation Effects in Fourier-Resolved Ptychography
G. Matteucci, G.F. Missiroli, G. POZZI, Univ. Bologna, Italy: Electron Holography and Electrostatic Fields
M. RADERMACHER, C. Lawrence, NYS Dept. Health, Albany, NY: Radon Transform Techniques for Alignment and Three-Dimensional Reconstruction from Random Projections
D. VAN DYCK, M. Op de Beeck, Univ. Antwerp, Belgium: From Image to Atomic Structure: How Far Are We?
M. VAN HEEL, E. Orlova, P. Dube, H. Stark, F. Zemlin, M. Schatz, Fritz Haber Inst., Berlin, Germany: Angular Reconstitution: High-Resolution Three-Dimensional Structure From Single Particles
E. VOELKL, L.F. Allard, Oak Ridge Natl. Lab., TN, B. Frost (Univ. Tennessee): Electron Holography: Recent Developments and Applications
H.S. von Harrach, VG Scientific, E. Grinstead, U.K.: Maximum Entropy Reconstruction of STEM Images
Z.L. Wang, Georgia Inst. Tech., Atlanta: Thermal Diffuse Scattering in Energy Filtered Electron Diffraction and Imaging
This conferences follows the Scanning Microscopy 1996 meeting in Bethesda, Maryland from May 11-16 (contact Om Johari at SMI for more information).
Papers can still be offered, please contact one of the organizers (see above).
Yves Maniette wrote: } } Dear all, } } I am looking for the fax number (or even E-Mail address) of the company } AREMCO products, OSSINING,NY. They sell Aremco crystalbond 509 glue, that } we use for preparing samples. } } thank you in advance } } Yves MANIETTE } Universitat de Barcelona } Serveis Cientifico Tecnics } Unitat ESCA TEM } Carrer Luis Sole i Sabaris } E-08028 BARCELONA } } Tel +34 3 402 16 95 } Fax +34 3 402 13 98
Aremco Products, Inc. P.O. Box 429 Ossining, NY 10562-0429 (914)762-0685 (914)762-1663 (FAX)
I do not have e-mail address.
--
Naresh Shah CFFLS, 341 Bowman Hall University of Kentucky Lexington, KY 40506-0059 e-mail: naresh-at-pop.uky.edu (606) 257-5119 (606) 257-4027 (Dept. office) (606) 257-7215 (FAX)
Crystalbond is available from AREMCO. Their phone is 914 762-0685 fax: 914 762-1663.
Joe Geller Geller Microanalytical Lab 426e Boston St. Topsfield, MA 01983-1212
On Wed, 24 Jan 1996, Yves Maniette wrote:
} } Dear all, } } I am looking for the fax number (or even E-Mail address) of the company } AREMCO products, OSSINING,NY. They sell Aremco crystalbond 509 glue, that } we use for preparing samples. } } thank you in advance } } } } Yves MANIETTE } Universitat de Barcelona } Serveis Cientifico Tecnics } Unitat ESCA TEM } Carrer Luis Sole i Sabaris } E-08028 BARCELONA } } Tel +34 3 402 16 95 } Fax +34 3 402 13 98 }
Those of you interested in callculations such as those involved in processing EDS spectra should by all means get a copy of the book "Data Reduction and Error Analysis for the Physical Sciences" by P. R. Bevington, McGraw-Hill, 1969 It contains about a dozen self-consistent FORTRAN programs for carrying out many of the basic mathematical operations involved in such calculations. W. C. Bigelow (bigelow-at-umich.edu)
} I'm trying to get in touch with TSL TexSEM Laboratories Incorporated a } company that makes EBSP software for Windows. I have a phone number } 801-467-9930, but nobody answers. Anyone know how to get in touch with } them?
I am looking for any manuals and/or technical data, i.e. circuit diagrams, on a Hitachi S-310A field emission SEM. We have the column and vacuum control/power supplies unit but no scan/display unit and I want to find out how to connect a computer-based scanning/image acquisition system in its place for a project.
I have been having a problem during "drop" staining of grids for TEM: I place my slot grid on a drop of stain. Sometimes the formvar which holds the sections in place detach from the grid - the formvar (and sections) float on the drop of stain and the grid sinks to the bottom.Has anyone out there been having a similar problem? Do you know of any solution. I have taken a number of "remedial" measures but I am not sure whether they have worked since the problem re-occurs from time to time. For example I have : - stained grids after allowing them to "stand" for at least 24 hours after sectioning. - tried multiple grid staining - used a glue pen - pre-dipped grids in formvar before picking up sections - cleaned grids thoroughly in acetone-HCl- DW -used acetone-stored grids -left grids with sections (on formvar plates) in a desiccator for extended periods I would appreciate any suggestions.
} One of my courses will discuss in length the contrast } theory used for the TEM. } Since I've never ever worked with these beasts I wonder if there is a } S/W simulator or some other means that wouldmake the digestion of the } theory a little easier ??? } } I have to start from electron diffraction, white field, dark field, } Kikuchi pattern etc... all new to me ! to get to the contrast theory.
Dear CFILION, The Academic Press volumes, Principles of Electron Optics, by Hawkes & Kasper, go into this in great detail in Vol. 3. Good luck. Yours, Bill Tivol
********** PLEASE RESPOND BY EMAIL TO:jbpawley-at-facstaff.wisc.edu: **********
LIMITED SPACE STILL AVAILABLE. ________________________________________________________________________________
Second announcement:
An intensive, 8-day course on
"3D MICROSCOPY OF LIVING CELLS"
will be given at the
University of British Columbia, Vancouver, BC, Canada
July 27 - August 4, 1996
THE PURPOSE OF THE COURSE
Modern methods of 3D light microscopy promise a revolutionary improvement in our ability to view living cells. To help convert this promise to reality for a wider selection of biological scientists, an intensive eight day residential course concentrating on all aspects of the 3D Microscopy of Living Cells will be introduced at the University of British Columbia, in the summer of 1996. Covering everything from basic microscopy to the technical considerations that define the highest levels of performance of the confocal microscope, this course will include:
* Quantitative confocal microscopy * Pixelation: The Nyquist Criterion * Lasers and laser tweezers * Objectives and aberrations * Scanning-systems * Wide field/deconvolution techniques * Detectors: operation and performance * Optimal pinhole size & photon efficiency * Dye design, characteristics and use * How to keep your cells alive * Two-photon excitation * Video-rate confocal imaging * Measuring ion concentrations * Display and measurement of 3D data * Digital hard copy and storage * Fluorescent & gold labeling of living cells * Backscattered light imaging.
Lecture demonstrations will be interspersed with hands-on laboratory exercises that will utilize all of the currently available commercial instruments for 3D microscopic imaging. Students will work in groups of three throughout the discussion and laboratory sessions, and will complete a live-cell 3D study on a specimen of their choice. At least seven, separate 3D microscopical workstations, attended by a technical staff of 15, will be available for student use. Overall, the teacher/student ratio will be more than 2:1.
International Academic Faculty:
* Jon Art University of Illinois * Milton Charlton University of Toronto * Rachel Errington Oxford University * Jim Pawley University of Wisconcon-Madison * Wallace Marshall U. of California, San Francisco * Ernst Stelzer EMBL, Heidelberg * Roger Tsien University of California, San Diego * Pavel Vesely Academy of Sciences, Prague * Watt Webb Cornell University * Michael Weis University of British Columbia * Nick White Oxford University
International Commercial Faculty:
* Ernst Keller Carl Zeiss, NY * Paul Millard Molecular Probes, OR * Sigrid Myrdal Bristol-Myers Squibb, WA * K. Sam Wells Bio-Rad,,CA
WHO SHOULD ATTEND?
The course is designed for biological research scientists and advanced graduate students who use, or plan to apply 3D microscopy to studies involving living cells. No previous experience in advanced light microscopy is required but applicants will be asked to outline how they plan to use 3D microscopy and describe a short research project that they plan to carry out during the course. Most projects will If space permits, students with other interests in 3D light microscopy will be welcomed.
PLAN OF INSTRUCTION
Classes will meet from 8:30 -12:30 and 1:30 - 5:30 and will alternate between lecture-demonstrations and laboratory sessions. From Monday to Friday, facilities and supervision will be available until 11:00 pm, for students to work on their projects. On average, only two topics will be covered in each morning or afternoon session. There will be enough 3D microscopy setups to permit students in groups of three, to "learn-by-doing" following each demonstration. Lab handouts will include detailed questions to stimulate group discussions. Prior to the course, students will be organized into groups and encouraged to communicate by email/phone, about the "Living-cell" group projects that they will pursue in the evenings and that will be presented to the class on the last day of the course. Students must contact the Course Organizer to make necessary arrangements for the transport and maintenance of cell lines etc. needed for their projects.
ACCOMMODATIONS
Campus accommodations border on the luxurious and arrangements can easily be made for accompanying family members. Rooms and suites will be situated in the Walter Gage Residence located centrally, a few blocks from either the lecture-lab facilities or the Student Union Building which contains a large cafeteria, lounge, bank, etc. Many of the rooms in the Gage Residence have breathtaking views of the mountains of North and West Vancouver, and the Pacific Ocean. A variety of accommodation types are available:
- Single room with shared washroom $24
- Single room with private bath $41
- Double suite (kitchenette, private bath, TV, phone, double bedroom plus separate sitting room) $63
- Triple suite (twin bedroom and queen Murphy bed in sitting room, balcony, kitchen, bath, TV, phone) $74 (All fees are $ US per night. Add 15% VAT)
Students are encouraged to bring family members to enjoy the pristine beauty of the Vancouver area and the miles of sandy beaches that surround the campus.
TUITION
Tuition is $1,500 US. On receipt of the deposit, all students will receive preliminary assignments and a copy of the textbook, "Handbook of Biological Confocal Microscopy," (Plenum, 1995). The tuition fee includes one ticket for the opening reception and the banquet, the textbook and all handouts. Accommodations and meals are not included in the tuition fee.
APPLICATIONS
Applicants will complete a questionnaire to assess knowledge level and field of interest. Enrolment will be limited to 20 participants. Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with basic texts to read before the course begins. Application packages may be obtained from
Prof. James Pawley, Rm. 1235, 1500 Johnson Dr., Madison, WI, USA 53706. Phone: 1-608-263-3147, Fax 1-608-265-5315, Email: jbpawley-at-facstaff.wisc.edu
Application deadlines:
Applications forms requesting information on field of interest and level of experience must be received for screening by March 1, 1996. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 1996. In general, refunds of the deposit will not be possible.
Applications must be received by Mar. 1/96 50% deposit due Apr. 15/96 Registration 3:00 - 5:00 pm Sat., July 27/96 Last class will end with lunch Sun., Aug. 4/96
*****************************************
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
Message-Id: {199601242132.PAA23101-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 08:55 AM 1/23/96 -1000, you wrote: } As I am in one of the facilities who used to have a "pre-paid, lump-sum" } system in place that appeared to make everybody happy, only to have a } federal audit come in and SEVERLY penalize us for doing so, I would also } be interested in what other recharge facilities are doing. Please keep } this dialog going, or post a compilation of replies! } } Mahalo, } Tina } ************************ Tina -
I am also in a fee-for-service situation. I get no support from grants, except that investigators use funds from their grants to pay me for the electron microscopy I do for their research projects. We have a fee structure that basically prices our services depending upon what we are called on to do, and whether there is a signed report that goes into a patient's file in clinical cases. Some time ago when I was setting this up and was curious what other service EM labs, especially medical labs like my own, charged for their work, I was told that asking other labs what they charged was illegal and amounted to price fixing. I am not sure what agency enforces that, probably Medicare; and I am not sure if it is still true. But if you have medical cases, you might inquire about this.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
About a year ago, after using EM's for 27 years and having developed cataracts in both eyes at an unusually early age (49), I asked the question through this medium about eye damage resulting from long-term use of electron microscopes. As in the current discussion some interesting comments were made. There is definitely evidence that radiation from a variety of sources causes cataracts, typically in the posterior capsule of the lens - not the nucleus where senile cataracts usually develop. However, modern electron microscopes should be adequately screened to prevent this. In my case the cataracts were apparently typical of radiation-induced cataracts although when and where the irradiation took place is anyone's guess. Someone has mentioned UV radiation from vacuum evaporators. This could be a possibility because I know of little in the way of precautions which are routinely taken to prevent this.
Robin Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za (tel: +27 461 318168 - fax: +27 461 24377)
} Dear Dr. Cheng, } } Thank you for your reply. } } I have not had good success with the E.mails publicity. I provide the } latest flier below. Can you please help me by having it posted? Any other } help you can give to publicize the conference will be most appreciated. } ----
-- [ From: Dr. Om Johari * EMC.Ver #2.0 ] --
Please Circulate
Scanning Microscopy International P.O. Box 66507, Chicago (AMF O'Hare), IL 60666, USA Telephone: (708) 529-6677 / FAX: (708) 980-6677 E.mail:73211.647-at-compuserve.com
Fifteenth Pfefferkorn Conference on Electron Image and Signal Processing
May 18-22, 1996 at Silver Bay Association, Silver Bay, NY
The Conference will provide a forum for discussion of the current status, recent advances and prospects of many aspects of image and signal processing in electron and near-field microscopy and microanalysis. The principal themes are three-dimensional reconstruction, image restoration, enhancement and analysis (including morphological and algebraic approaches in particular), electron holography, the phase problem, processing image sets (from STEM detectors and from defocus and tilt series, for example), image simulation, and hardware and software for image acquisition and processing and for microscope control. Please see below for a list of speakers.
The organizers are: Dr. Peter W. Hawkes, CNRS Lab. Opt. Elect., B.P. 4347, 31055 Toulouse Cedex, France (Phone: 33-62-257884 / FAX 33-62-257999 / E.mail: hawkes-at-cict.fr); Dr. W. Owen Saxton, Univ. Cambridge, U.K. (FAX: 44-1223-334 567 / E.mail: wos1-at-cus.cam.ac.uk); and Dr. Joachim Frank, NYS Dept. Health, Wadsworth Center, Albany, NY 12201-0509, USA (Phone: 518 474 7002 / FAX: 518 474 8590 / E.mail: joachim-at-wadsworth.org). Interested contributors should contact either of the organizers with a title and a one page summary; please also include other relevant information (e.g., time desired, reprints of relevant publications, etc.).
Full-length papers will be published in the Conference Proceedings to be issued as Scanning Microscopy Supplement 11, 1997.
Conference Information: Registration begins at 6 PM on Friday, May 17. The first lecture will be at 8:30 AM on Saturday, May 18; the Conference will end at lunch on Wednesday, May 22. Sessions will be daily in mornings (from 8:30 to 1), afternoons (about 4:30-7), and evenings (about 8:45 to 10). Ample discussion time will be allowed; active participation by attendees is invited. Because of the limited number of attendees, the Pfefferkorn Conferences provide an excellent opportunity for in-depth discussions and personal contacts. Conference attendance is by application or invitation only. Qualified registrants will be accepted on a first-come-first-served basis. To apply: please submit name, mailing and E.mail addresses, phone (work and home) and FAX numbers, and a 50-100 statement about your qualification relative to the theme (and topics) of the conference, accompanied by the full registration fee of US $200 (includes attendance to sessions, coffee-breaks, and a copy of the Conference Proceedings).
Conference Location: The sessions will take place at one of the conference rooms at Silver Bay Association (SBA), Silver Bay, NY 12874 (phone: 518 543 8833 / FAX: 518 543 6733 / E.mail: sbaconf-at-aol.com), located 80 miles north of Albany, NY. This 600 acres beautiful, peaceful, registered historical place, located on Lake George and in the Adirondack mountains, offers dozens of water and land sports, gym programs, hiking, etc. and is filled with gardens, rolling lawns and gentle streams. It is ideal for a family vacation or retreat. The room rates (all rates include three full meals daily!) range from $40 per person (in two and four bed-room cottages ideal for four and eight persons, respectively), to $55 and $65 per person (based on two persons to a room) in rooms without and with private baths, respectively. Accompanying children aged 4 to 12 pay half of these rates. Single rooms are available at a cost of $69 (without private bath) or $80 (with private bath). It is expected that all attendees will stay at this location. To make accommodation reservations, please contact SBA directly and inform us of your travel details (mode and day/time). Details about getting to Silver Bay will be sent to all attendees: nearest airports are Albany (New York) and Burlington (Vermont); foreigners may find it easier to fly to Montreal, Canada (about 150 miles north) or New York (about 200 miles south). There is one daily train each way from New York and Montreal which stops in Ticonderoga, NY (about 20 miles from Silver Bay); some bus service is available too. Appropriate van transportation from Albany (at costs ranging from $50 per van for less than 4 people, or $10 per person for more than 5 persons) will also be arranged by SBA. Attemps will also be made to form car pools with those driving.
Past Conferences: Started in 1982, in honor of the late Prof. Gerhard E. Pfefferkorn, these Conferences present an in-depth discussion of fundamental topics in scanning microscopy and related techniques. Two conferences on topics directly related to the theme of the 1996 Conference were held in 1987 and 1991; registrants to the 1996 Conference can obtain those proceedings [containing original peer-reviewed, full-length papers published as Scanning Microscopy Supplements 2 (1998) and 6 (1992)] at special package prices (including uninsured cheapest rate mail delivery) of $95 (for US delivery) and $105 (for outside US delivery).
Scanning Microscopy International (SMI), a not-for-profit organization, sponsors the annual Scanning Microscopy, Cells and Materials, and Food Structure meetings: 1996 meeting will be from May 11 to 16 at Hyatt Regency Hotel, Bethesda, Maryland (suburb of Washington, D.C.]; publishes the international journals: Scanning Microscopy (previously Scanning Electron Microscopy till 1986), Food Structure, Cells and Materials (from 1991), and Scanning Microscopy Supplements (Proceedings of the Pfefferkorn Conferences); and issues publications devoted to selected topics derived from its journals. Details of the programs planned for the Scanning Microscopy 1996 meeting, a complete current list of publications, samples Tables of Contents of publications, etc. are available on request. The topics of this 1996 Conference are relevant to the themes of our journals; papers for publication are invited (Instructions for Authors are included in our journals or available on request).
For more information, please contact Dr. Om Johari, at SMI.
---
List of speakers as of January 22, 1996 (note: * = invited but not formally accepted as yet; when there are multiple authors, speakers name is in CAPITALS).
G. ADE, Physikal.-Techns. Bundes., Braunschweig, Germany: (1) A Digital Method for Noise Reduction in Holographic Reconstructions and Electron Microscopical Images (2) Coma-Free Alignment of Electron Microscopes and Digital Determination of Aberration Coefficients (with R. Lauer)
G. Anstis, Univ. Technol., Sydney, Australia; C.R. Birkeland, S.C. Anderson, D.J.H. Cockayne: Computation and Quantitative Analysis of HAADF Lattice Images of GaAs
N. Boisset, J.C. Taveau, V. You, F. de Haas, J. Lamy, URA CNRS, Tours, France: Refinement of Single Particle 3D Reconstruction (Art Method) Based on Topological Selection of EM Views
N. Bonnet, Univ. Reims, France: Multi-Dimensional Spectrum and Image Processing
C.B. Boothroyd, Univ. Cambridge, U.K.: Recent Results on Energy Filtered Image Quantification
C. BURMESTER, Max-Planck-Inst. Mol. Physiol., Dortmund, Germany; K.C. Holmes, R.R. Schroeder (Max-Planck-Inst. Med. Forschung, Heidelburg, Germany): Feasibility of the Multiple Isomorphous Replacement Method in Protein-Electron-Diffraction
H. CHENG, T. Baker, Purdue Univ., W. Lafayette, IN: The Correlation Approach to Virus Phasing
*M. Coster, Univ. Caen, France: Morphology and SEM Image Analysis: Recent Progress
C. DINGES, H. Rose, Tech. Hochsch., Darmstadt, Germany: Simulation of TEM Images and Diffraction Patterns Considering Phonon and Electronic Excitations
J. Frank, NYS Dept. Health, Albany, NY: Three-Dimensional Reconstruction (exact title to come)
S. Swaminathan, I.P. Jones (Univ. Birmingham, UK), N.J. Zaluzec (Argonne Natl. Lab.), D.M. Maher (No. Carolina State Univ., Raleigh), H.L. FRASER, Ohio State Univ., Columbus: Determination of Accurate Low Order Structure Factors for Si and TiAl Using Energy Filtered CBED
D.R. Beniac, G.J. Czarnota(1), F.P. Ottensmeyer(1), G. HARAUZ, Univ. Guelph, Canada; (1 = Ontario Cancer Institute, Toronto): Challenges of Three Dimensional Reconstruction of Nucleoprotein Complexes from Electron Spectroscopic Images
P. Kruit, B.M. Mertens, Delft Univ. Technol., Netherlands: Image Acquisition by Scanning in Fourier Space
M.K. KUNDMANN, Gatan EELS Software, Downers Grove, IL; S.L. Friedman, A.J. Gubbens, O.L. Krivanek: Characterization and Correction of TEM Energy Filter Aberrations
P.L. Bellon, S. LANZAVECCHIA, Univ. Milan, Italy: The Moving Window Shannon Reconstruction in Real and Fourier Domain. Applications in Tomography
H. Lichte, Univ. Dresden, Germany: Pitfalls on the Road to 0.1 nm with Electron Holography
M. MANKOS, IBM Watson Res. Ctr., Yorktown Hts., NY; M.R. Scheinfein, J.M. Cowley (Arizona State Univ.): Holography and the STEM
M.R. McCartney, Arizona State Univ., Tempe: Recent Applications of Electron Holography
D.G. MORGAN, D.J. DeRosier, Brandeis Univ., Waltham, MA: Analysis of Disordered Helices Using Correlation Methods
P.D. NELLIST, S.J. Pennycook, Oak Ridge Natl. Lab., TN: Probe and Object Function Reconstruction in Incoherent STEM Imaging
M.A. O'Keefe, Lawrence Berkeley Lab., Univ. Calif., Berkeley: On-Line Remote-Control Electron Microscopy with Emphasis on the Image and Signal Processing
P.A. PENCZEK, J. Zhu, R. Schroeder, J. Frank, NYS Dept. Health, Albany, NY: Three-Dimensional Reconstruction with CTF Compensation from Focus Series
T. Plamann, Univ. Bristol, U.K.: Three-Dimensional Propagation Effects in Fourier-Resolved Ptychography
G. Matteucci, G.F. Missiroli, G. POZZI, Univ. Bologna, Italy: Electron Holography and Electrostatic Fields
M. RADERMACHER, C. Lawrence, NYS Dept. Health, Albany, NY: Radon Transform Techniques for Alignment and Three-Dimensional Reconstruction from Random Projections
D. VAN DYCK, M. Op de Beeck, Univ. Antwerp, Belgium: From Image to Atomic Structure: How Far Are We?
M. VAN HEEL, E. Orlova, P. Dube, H. Stark, F. Zemlin, M. Schatz, Fritz Haber Inst., Berlin, Germany: Angular Reconstitution: High-Resolution Three-Dimensional Structure From Single Particles
E. VOELKL, L.F. Allard, Oak Ridge Natl. Lab., TN, B. Frost (Univ. Tennessee): Electron Holography: Recent Developments and Applications
H.S. von Harrach, VG Scientific, E. Grinstead, U.K.: Maximum Entropy Reconstruction of STEM Images
Z.L. Wang, Georgia Inst. Tech., Atlanta: Thermal Diffuse Scattering in Energy Filtered Electron Diffraction and Imaging
This conferences follows the Scanning Microscopy 1996 meeting in Bethesda, Maryland from May 11-16 (contact Om Johari at SMI for more information).
Papers can still be offered, please contact one of the organizers (see above).
Message-Id: {9601242300.AA00507-at-zell.enk.ks.se} X-Sender: martin-at-136.155.21.21 X-Mailer: Windows Eudora Light Version 1.5.2 Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable X-Priority: 2 (High)
Equipment for "UNCAGING OF CAGED SUBSTANCES"?
I wonder what equipment is used or should be used for uncaging (photolysis) of caged (photoactivable) compounds.
Background: Caged compounds usually seems to be optimally activated by a short (millisecond) pulse of {365 nm light. To get this light one may use light sources like lasers, flash lamps or maybe arclamps with fast shutter. The light may reach the sample either through the objective (light through epifluorescence port of the microscope) or by leading an optical fibre directly to the cell.
Questions about instrumentation:
- What light sources are used (types of lasers, Xenon arc lamps, Mercury arc lamps, flash lamps)?
- How is the exact light path to the cell/cells/subcells? What types of optics and components are used?
- What effect or energy is needed?
- What companies provide necessary parts (like Oriel, Newport, Melles Griot, Hamamatsu)?=20
- What complete commersial systems are available for this, and how are their performance?
Thanks in beforehand for your answers, Martin
----------------------------------------------- Martin K=F6hler Department of Molecular Medicine Rolf Luft Center for Diabetes Research L6B:1 Karolinska Hospital S-171 76 STOCKHOLM SWEDEN phone 46-8-7295732 46-8-7295725 fax 46-8-7293658 E-mail mk-at-enk.ks.se ---------------------------------------------
Here is another message about glues for sample preparation. Hope it helps. YM.
I saw your message om the Crystalbond. Please note that we sell the Crystalbond in our catalog and at very competitive prices. If you are interested please see page 147 of catalog XII. If you do not have this catalog please let me know and we will send you one. I hope this information helps.
Stacie Kirsch Electron Microscopy Sciences. Tel: 215-646-1566 Fax: 215-646-8931
Just a note of thanks to all those who provided their views and recommendations regarding the use of ultramicrotome quality glass (UMG) v's histology quality glass. The upshot is that UMG should be more than adequate for the purpose of making Ralph knives. Thanks again for your interest.
Brett. Brett W. Cockman Technologist in Charge School of Dentistry University of Western Australia Voice: (619-2205834) Fax: (619-2213829) e-mail; bcockman-at-uniwa.uwa.edu.au
I got an interesting piece of information about glues for TEM sample preparation, from D. Henriks, South Bay Technology. So I am sending it to the list, as it might be of interest. Particularly I know that it is quite difficult to get the AREMCO 509 glue in Europe, because it has to go through a network of distributors and sub-distributors, so that the price comes to be quite elevated.
-- MESSAGE --
We can supply it to you in the same 7" long sticks that you get from Aremco or you can buy a package of 20 3" long x 1/4" x 1/4" Unwrapped sticks. These are ideal for sample preparation. I absolutely guarantee that you will be 100% satisfied with the product.
Below, I have listed quantity pricing and alternative packaging for the QuickStick 135. This material is the same as "Crystalbond 509".
20 x 3.5" Sticks per tray (360 grams per tray) - this is our standard package. Minimum order is 1 tray.
1 x .875" diameter x 7" long wrapped in cardboard tube (5 sticks = 450 grams). Minimum order is 10 sticks.
10 x .875" diameter x 7" long unwrapped in plastic tray (1 tray = 900 grams). Minimum order is 1 tray.
1 pound bulk pack trays Minimum order is 10 pounds
Sizes and gross weights for above are approximate.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com sbt-at-msa.microscopy.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
Detachment of formvar films from grids during staining is a common problem. We recommend use of the SynapTek grid stick kit, which holds the grids during staining in the same plane as the solution flow, minimizing the chances of losing the films (or collecting surface debris). The grids are held to the grid stick with a special adhesive which is not attacked by solvents, and which will not remain on the grid once it is removed from the stick. Please contact me directly for specific ordering information. Steven Slap, Vice-President Energy Beam Sciences, Inc 75767,640-at-compuserve.com
I used the Crystalbond 509 glue forTEM prep initially, but about a 2 years ago I switched to quick-bond nail glue (for cosmetic fingernails - a cyanoacrylate formula, like SuperGlue) for several reasons: 1. specimens do not have to be heated, which seemed to be a problem with the specimen mounts for the dimpler I was using
2. the glue thickness is about half of what I could achieve with Crystalbond, which increased my success rate at preparing copper-sapphire interface specimens
3. specimen adhesion is nearly instantaneous - instant gratification!
4. I can walk across the street and purchase more instead of waiting 3 weeks for the purchase order to get through purchasing
5. Crystalbond properties seemed to degrade during our humid summers.
The only real problems with the stuff are that it takes ~2 hours of soaking in acetone to remove specimens from the dimpler mount, and when not capped tightly an entire bottle will cure in about 2 weeks.
For larger jobs, I use a homemade version of the Crystalbond glue (found in an old Handbook of Chemistry and Physics) which can be made by mixing equimolal quantities of phthalic anhydride and ethylene glycol for 24 hours at 200 deg. C in a covered beaker, stirring every couple of hours. Pour into some sort of flexible mold to make shapes and store in a cool dry place.
-Kirk _____________________________________________ Kirk Rogers krogers-at-materials.ecn.purdue.edu OR kirk.a.rogers.1-at-purdue.edu Purdue University, School of Materials Science and Engineering, 1289 MSEE building, W. Lafayette, IN 47907-1289 OFFICE: 317-494-8751 FAX: 317-494-1204 http://materials.ecn.purdue.edu/~krogers
For planktonresearch I use an old Leitz Ortholux stand.On this stand a socalled Berek condensor is used with interchangeable toplenses. I look for an small toplens of NA 1.4 which fits onto that condensor. Can any microscopist help me , for a reasonable price.
Thank you in advance
Pieter Houpt
Pieter.M.Houpt. Marine Plankton & microscopy tel/fax. 003170504466 The Hague. The Netherlands.
Message-Id: {199601251627.KAA22140-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 08:51 AM 1/25/96 -0600, you wrote:
} Let me get this straight: you are required to charge "customary fees" for } your services (otherwise you won't get paid), but you are not allowed to } find out what the usual fees are? Did anyone ever try a class in elementary } logic on these guys who make the rules? } ******************* I just went down the hall to check with our administrative associate in charge of billing. Yes, I am right: for me (Director of the medical EM lab) to ask people in another medical lab, not within our organization, what their charges are for their services is illegal. As Chuck Garber, points out, it runs afoul of the anti trust laws. I can ask for, and the other lab can give me, a RANGE within which their charges fall, but they cannot disclose, nor can I demand to know, what the actual numbers are. No, I do not know how broad that "range" must be.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
Message-Id: {199601251627.KAA22136-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 07:22 AM 1/25/96 EST, you wrote:
} While you should not take this as legal advice, at no time has the } congress ever exempted nonprofit or other tax exempt organizations from } the anti trust laws. If anyone should ever challenge you on that one, } ask them to show you where nonprofits were ever given any such } exemption. ******************* I assumed that to be the case. *******************
} Secondly, I for one, sitting in the private sector, get quite a kick } out of these kinds of conversations. It sounds to me like the decision } on what to charge someone is about as arbitrary as could be, and there } also seems to be an admission that there is little relationship between } what someone costs vs. how it is charged. ******************* For years that's exactly what I based my charges on, ie. my costs plus a bit to cover replacement and depreciation. The goal was to run a lab that paid for itself. There was no pressure to make a profit strictly for profit's sake. Now with the new politics in health care, the "bottom line" is more important than patient care, and I am told what to charge. *******************
} Yet, how would you feel if I was selling you a sputter coater 20% more } than I would charge someone else only because, say, I thought you could } afford it more than someone else? I am sure you would not feel very } good about that at all, as indeed you should feel.
******************* Hey, it happens! Not from reputable suppliers such as SPI....but it happens.
I appreciate your thoughts on these matters, Chuck, because you view things from a different perspective.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
I have been asked to image the interior structure of a polymer suspension.
Ideally I would like to resin embed for TEM. I am not a polymer scientist, but I do know that these polymers produce spheres when suspended with small amounts of BSA. The polymer is polyglycolic acid (PGLA). I called the manufacturers, but they have not done any significant imaging of it, and do not appear to be interested. They did tell me that the PGLA is heat sensitive and dissolves in isopropal alcohol. My experiment with epoxys was an unmitigated disaster, and I am considering a low temperature embedment procedure (Another researcher tried Lowicryl, but the spheres did not remain intact).
I referenced a book entitled "Polymer Microscopy" by Sawyer and Grubb, but it doesn't refer to anything similar to my polymer.
If this question has been asked before, I apologize. I need information on your experiences with light microscopy CCD camera systems. What systems are available for easily and inexpensively (?) digitizing images from light microscopes to PC formats? I need manufacturers (addresses) and prices please. Please send information to my e-mail address--especially if this discussion has taken place before. Thank you.
John C. Wheatley Lab Manager Center for High Resolution Electron Microscopy BOX 871704 Tempe, AZ 85287-1704 Phone: (602) 965-3831 FAX: (602) 965-9004 John.Wheatley-at-ASU.Edu
Alternate-Recipient: allowed Auto-Forwarded: prohibited Content-Return: allowed Disclose-Recipients: prohibited Conversion: allowed Importance: normal Priority: normal Sensitivity: Company-Confidential
} I wonder what equipment is used or should be used for uncaging (photolysis) } of caged (photoactivable) compounds. } [snip] } - What light sources are used (types of lasers, Xenon arc lamps, Mercury arc } lamps, flash lamps)? } } - How is the exact light path to the cell/cells/subcells? What types of } optics and components are used? } [snip]
Dear Martin and especially SPM'ers on this list, Can this be done using either a NSOM probe or an NSOM-like input probe? If so, this would likely have many uses. Yours, Bill Tivol
CALL FOR PAPERS (December 1995) (Deadline for Submission of Abstracts is April 1, 1996)
=09COMBINED MEETING =0945th ANNUAL DENVER X-RAY CONFERENCE =09=09- and - =09POWDER DIFFRACTION SATELLITE MEETING OF THE XVII=20 =09CONGRESS OF THE INTERNATIONAL UNION OF CRYSTALLOGRAPHY
August 3-8, 1996, Denver, Colorado, U.S.A.
Co-Sponsored by the IUCr; the ICDD; and the University of Denver=20 Department of Engineering=20
=09The meeting will be held August 3-8, 1996, at the=20 Marriott Denver Tech Center Hotel,=20 4900 S. Syracuse St., Denver, CO 80237, U.S.A. =20 Phone (303) 779-1100, Fax (303) 740-2523. =20
Prospective attendees should make reservations early. The number of=20 rooms at the special conference rate is limited. The Conference rate=20 of $76.00 per day plus 11.8% tax for a single or double room can be=20 obtained if reservations are made before July 1, 1996, subject to=20 availability. Also available are a limited number of rooms for=20 students at $22.50 per night (double occupancy).=20
The Conference has appointed Monarch Travel as the official travel=20 agency to help with travel and car rental arrangements. Please contact=20 Monarch Travel by phone at (800) 458-7177, or (215) 557-7177=20 outside the U.S., or fax (215) 563-0479, with travel questions. The=20 program is tentatively as follows:
PLENARY SESSION =20 =09GRAZING-INCIDENCE X-RAY ANALYSIS Organizers:=09T. C. Huang, IBM Almaden Research Center, San Jose, CA=20 =09=09R. J. Cernik, Daresbury Lab, Warrington, England Opening remarks: by P. K. Predecki, University of Denver Presentation of the 1996 Birks Award: to John V. Gilfrich, SFA,=20 =09Inc./NRL, Washington, DC Presentation of ICDD Distinguished Fellow Awards: to J. W. Visser,=20 =09Delft, Netherlands; and D. K. Smith, =09Pennsylvania State University
INVITED PAPERS: "Grazing Incidence Small-Angle X-Ray Scattering," J. B. Cohen,=20 =09Northwestern Univ., Evanston, IL
"Depth Scaling of Phases in Thin Films," H. Goebel, Siemens AG=20 =09Corp., Munich, Germany=20
"Grazing-Incidence Technique for Surface, Interface, and Thin Film=20 =09Analysis," T. C. Huang,=20 =09IBM Almaden Research Lab, San Jose, CA; and=20 =09P. K. Predecki, Univ. of Denver, CO=20
"Grazing Angle X-Ray Spectrochemical Analysis, Present and=20 =09Future," Y. Gohshi, Univ. of Tokyo=20
"Analysis of Layered Materials Using Glancing-Incidence X-Ray=20 =09Reflection," D.K.G. de Boer and =09 =09A.J.G. Leenaers, Philips Research Labs, Eindhoven,=20 =09The Netherlands
SPECIAL SESSIONS=20
- PHASE QUANTIFICATION (CPD sponsored) Organizers: D. L. Bish, Los Alamos National Lab, NM (505) 667-1165,=20 =09=09fax (505) 665-3285 =09=09D. K. Smith, Pennsylvania State University, PA (814) 865-5782,=20 =09=09fax (814) 863-7845
INVITED PAPERS: "Quantitative Phase Analysis Using the Full Powder Diffraction=20 =09Pattern - Its Development and Current Status,"=20 =09R. J. Hill, CSIRO, Port Melbourne, Australia
- THIN FILMS AND MULTILAYERS (CPD sponsored) Organizers: H. Goebel, Siemens AG, Germany fax 49 89 636 42256, =20 =09=09herb.goebel-at-zfe.siemens.de =09=09D. E. Cox, Brookhaven National Lab, NY (516) 282-3818,=20 =09=09fax (516) 282-2739
INVITED PAPERS: (for Thin Films and Multilayers session) "The Domain Structure of Langmuir-Blodgett Multilayers," U.=20 =09Pietsch, Univ. of Potsdam, Germany "X-Ray Analysis of Magnetic Multilayers," V. Valvoda, Charles=20 =09Univ., Prague, Czech Republic "Quantitative Characterization of Textures in Thin Films:=20 =09Measurements and Analysis," H. R. Wenk,=20 =09Univ. of California, Berkeley
INVITED PAPERS: "Routine X-Ray Fluorescence Spectrometric Analysis by VERBA- =09XRF," P. Verkhovodov, Kiev, Ukraine "The Permutations and Combinations of Quantitative XRF=20 =09Methods," G. Lachance, Hammond, Ont., Canada "An Analysis of Secondary Enhancement Effects in Quantitative=20 =09XRFA," M. Mantler, Technical Univ. of Vienna
- TOTAL REFLECTION XRF Organizers: M. A. Zaitz, IBM East Fishkill, NY (914) 894-6337,=20 =09fax (914) 892-6256 =20 =09=09A. Shimazaki, Toshiba Corp.
INVITED PAPERS: "A Review on Principles and Recent Developments in TXRF," H.=20 =09Aiginger, Atominstitut, Vienna, Austria "New Possibilities and Applications of Total Reflection X-Ray=20 =09Fluorescence Analysis," A. Knochel,=20 =09University of Hamburg, Germany
- STRAIN/STRESS DETERMINATION BY DIFFRACTION METHODS Organizers:=09A. Winholtz, University of Missouri (314) 882-6322,=20 =09=09fax (314) 884-5090 =09=09T. Ericsson, University of Linkoping, Sweden fax 46 1328 2505
INVITED PAPERS: "Dependence of the Elastic Constants for X-Ray Diffraction on the=20 Diffraction Plane,"=09I. C. Noyan,=20 =09IBM, Yorktown Heights, NY; and C. C. Goldsmith,=20 =09IBM East Fishkill Facility, NY. "Partially Destructive and Non-Destructive Determination of=20 =09Residual Stress States with Steep Subsurface Gradients," =09B. Eigenmann, Univ. Karlsruhe, Germany "Second Order Stresses in Single Phase and Multiphase Materials:=20 =09Examples of Experimental and Modeling =09Appproaches," J. L.=20 =09Lebrun, ENSAM, Paris, France=20
- DIFFRACTION PEAK PROFILE ANALYSIS (CPD sponsored) Organizers:=09R. L. Snyder, Alfred University, NY (607) 871-2438,=20 =09=09fax (607) 871-2392 =09=09D. Lou=EBr, University of Rennes, France =20 =09=09fax 33 9938 3487, louer-at-cicb.fr
INVITED PAPERS: "Model-Based Interpretation of Diffraction Line Profiles: The Case=20 =09of Lattice Distortion," E. Mittemeijer,=20 =09Technical University of Delft, The Netherlands "Model-Based Interpretation of Diffraction Line Profiles:=20 =09Determination of Crystallite Size and Shape,"=20 =09I. Langford, University of Birmingham, U.K. "Model-Based Interpretation of Diffraction Line Profiles: Strain=20 =09Broadening Caused by Dislocations," T. Ungar, =09Eotvos=20 =09University, Budapest, Hungary
- ENVIRONMENTAL APPLICATIONS OF XRF Organizers:=09R. Jenkins, ICDD, PA (610) 325-9810, fax (610) 325-9823 =09=09Y. Gohshi, University of Tokyo, Japan fax 81 3 3812 9254
INVITED PAPERS: "Use of XRF for Analysis of Contaminants in Seawater," T. Elam,=20 =09NRL, Washington, DC "Use of Semi-Quantitate Analysis Programs in the Analysis of=20 =09Pollutants," J. Croke, Philips Electronic=20 =09Instruments, Mahwah, NJ (Title to be announced) Y. Gohshi, University of Tokyo, Japan
- NEW DEVELOPMENTS IN DETECTORS AND OTHER X-RAY INSTRUMENTATION =09 =09(CPD sponsored) =09Organizers: H. Toraya, Nagoya Inst. of Technology, Japan =20 =09fax 81-572-27-6812, toraya-at-crl.nitech.ac.jp =09=09J. V. Gilfrich, SFA, Inc./NRL, Washington, DC (301)=20 =09=09365-5070 fax & phone
INVITED PAPERS: (for New Developments in Detectors session) =09"Novel Use of Imaging Plates in Powder Diffraction on the=20 =09Australian National Beamline Facility," D. Cookson, =09 =09Australian Nuclear Science and Technology=20 =09 =09"High Energy Resolution X-Ray Detectors Using Superconductors,"=20 =09D. Van Vechten =09 =09"Multiple-Detector System for Powder Diffraction with Synchrotron=20 =09Radiation," H. Toraya,=20 =09Nagoya Inst. of Technology, Japan.
- PRECISION AND ACCURACY IN STRUCTURE REFINEMENT FROM POWDER=20 DATA (CPD sponsored) Organizers:=09W.I.F. David, Rutherford-Appleton Lab, Chilton,=20 =09=09U.K. wifd-at-isise.rl.ac.uk =20 =09 R. A. Young, Georgia Inst. of Technology, Atlanta =20 =09=09(404) 894-5208, r.young-at-physics.gatech.edu=20
INVITED PAPERS: "Optimised Data Collection and Refinement Strategies for Powder=20 =09Data Analysis," W.I.F. David,=20 =09Rutherford-Appleton Lab, Chilton, U.K. =09"Structure Refinement with Combined X-Ray and Neutron=20 =09Diffraction Data - Successes and Failures,"=20 =09R. B. Von Dreele, Los Alamos National Lab, NM=20
- XRF ANALYSIS OF SEMICONDUCTOR WAFERS (BPSG) =09Organizers: R. Wilson, Rigaku/USA, Danvers, MA (508) 777- =092446 x124, fax (508) 777-3594 =09=09A. Iida, Photon Factory, Ibaraki, Japan =20 =09=09aiida-at-kekvax.kek.jp
INVITED PAPERS: "XRF Analysis of Thin Films and Problem Solving in the=20 =09Semiconductor Industry," M. A. Zaitz,=20 =09IBM, East Fishkill, NY. "Measurement Capabilities of X-Ray Fluorescence for BPSG=20 =09Films," J. Westphall,=20 =09Watkins-Johnson Company, Scotts Valley, CA "XRF Analysis of Insulator Films on Si Substrate," T. Konishi,=20 =09Asahi Chemical Industry Co., Japan
- HIGH-TEMPERATURE AND NON-AMBIENT APPLICATIONS OF DIFFRACTION Organizer: C. R. Hubbard, Oak Ridge National Lab, TN =20 =09(423) 574-4472, hubbardcr-at-ornl.gov
INVITED PAPERS: (To be announced)
TUTORIAL WORKSHOPS
- CREATING WINDOWING INTERFACES FOR FORTRAN=20 =09PROGRAMS: NEW LIVES FOR OLD TOOLS (1/2 day) =09B. Toby, NIST, Gaithersburg, MD; and L. Finger,=20 =09Geophysical Laboratory, Washington, DC
- TOTAL REFLECTION XRF (one day) =09P. Wobrauschek, Atominstitut, Vienna
- QUANTITATIVE TECHNIQUES AND ERRORS IN XRF (1/2 day) =09J. A. Anzelmo, Fisons Instruments
- QUANTITATIVE TECHNIQUES AND ERRORS IN XRPD (1/2 day) =09R. L. Snyder, Alfred University, NY; and R. Hill, CSIRO,=20 =09Australia
- NEW TECHNIQUES IN SEARCH-MATCHING (ICDD sponsored) (1/2 day) =09R. Jenkins, ICDD, PA; and D. K. Smith, Penn State=20 =09University
- E-MAIL SYSTEMS AND WWW RESOURCES (x-ray related) (1/2 day) =09R. Jenkins, ICDD, PA; and L. Cranswick, CSIRO, Australia
- SPECIMEN PREPARATION FOR XRF (one day) =09V. E. Buhrke, The Buhrke Company, Portola Valley, CA
- ANALYSIS OF METALS BY XRF (1/2 day) =09F. Feret, Alcan International, Canada
- XRD STANDARDS, CALIBRATION AND ALIGNMENT (1/2 day) =09J. P. Cline, NIST, Gaithersburg, MD; and R. W. Cheary,=20 =09Univ. of Technology, Sydney, Australia
- TEXTURE AND RELATED 3-D POLYCRYSTAL ANALYSIS (1/2 day) =09H. Bunge, Technical Univ. Clausthal, Germany; and R. Von=20 =09Dreele, Los Alamos National Lab, NM
- GRAZING INCIDENCE X-RAY CHARACTERIZATION (1/2 day) =09D. K. Bowen, University of Warwick, U.K.
- MICROANALYSIS APPLICATIONS XRD AND XRF (1/2 day) =09M. O. Eatough, Sandia National Labs, NM; and J. Brown,=20 =09University of Western Ontario, Canada
- STRATEGY FOR CALIBRATION OF XRF SPECTROMETERS (1/2 day) =09B. Vrebos, Philips, The Netherlands; and P. A. Pella, NIST,=20 =09Gaithersburg, MD
- METHODS OF CORRECTING FOR MATRIX EFFECTS IN XRF (1/2 day) =09R. Rousseau, Geological Survey of Canada
- SYNCHROTRON RADIATION ANALYSIS (1/2 day) =09P. Stephens, State University of New York; and A. Fitch,=20 =09ESRF, Grenoble, France =20 =09Contributed papers are hereby solicited for any of the above=20 special sessions or the XRD and XRF general sessions. Many of the=20 contributed papers will be placed in poster sessions. Those of more=20 general interest will be placed in oral sessions. =20
=09THE DEADLINE FOR SUBMISSION OF ABSTRACTS IS APRIL 1, 1996. =20
Abstracts are reproduced as-is in the abstracts book. They must not=20 exceed one page in length and must include title, author(s),=20 affiliation(s) and the text. They should be typed single-spaced using a=20 laser printer or equivalent with characters at least 2 mm high (lower=20 case) on a 15 cm wide x 20 cm long image area centered on an 8-1/2"=20 x 11" or A4 page, mailed flat without folds. On a SEPARATE=20 PAGE state: (1) speaker's name, mailing address, phone, fax and e- mail numbers; and (2) whether you intend to publish this paper in the=20 conference proceedings, Advances in X-Ray Analysis, Vol. 40. The=20 original and one copy of the abstract should be sent to:
=09Paul K. Predecki, Dept. of Engineering, University of=20 Denver, Denver, CO 80208, USA. =20
Additional information may be obtained from Lynne Bonno at the=20 above address, telephone (303) 871-3515, fax (303) 871-4450, or e- mail denxrcon-at-du.edu. A program for the Conference will be=20 prepared and mailed in May 1996. The program as well as this call for=20 papers will be placed on the University of Denver home page: =09http://littlebird.engr.du.edu/home.html =20
=09The Organizing Committee considers unprofessional the=20 withdrawal of a paper (except in special circum-stances) after it has=20 been accepted and widely advertised. Non-U.S. authors, in particular,=20 please try to secure travel funding and approvals before submitting=20 your abstract(s).
=09With regard to publication of presented papers, manuscripts=20 must be submitted no later than one month after the conference to=20 allow publication before the next conference. To be acceptable for=20 publication, papers should describe either new methods, theory and=20 applications, improvements in methods or instrumentation, or other=20 advances in the state of the art. Papers emphasizing commercial=20 aspects are discouraged. Information for preparing manuscripts will=20 be mailed in June 1996.
At 9:13 AM 1/24/96, Griffin, Robin wrote: } I'm trying to get in touch with TSL TexSEM Laboratories Incorporated a } company that makes EBSP software for Windows. I have a phone number } 801-467-9930, but nobody answers. Anyone know how to get in touch with } them?
Didnt they get bought out by Noran?
I heard a rumor.....
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu, John.F.Mansfield-at-umich.edu or jfmjfm-at-umich.edu they all reach me! URL: http://wwwpersonal.engin.umich.edu/~jfmjfm/jfmjfm.html
I recharge for use of specialized optical microscopes. Your University should have a budget analyst in Budget that can help you. You need to estimate the amount of non-use you expect and build that into your rate as an expense. You also charge for your overhead, salaries (figure in vacation & sick leave, breaks, etc.) and supplies. You also should calculate depreciation for your equipment and build a reserve so you can buy new equipment. Our campus even has a high-value equipment reserve that provides for catastrophic loss of an expensive scope. When you have figured all your costs, you should be able to divide it up as fits your program. For instance, charge a one-time lump sum for each use that recovers costs incurred regardless of the length of use, i.e. cleaning, prep of scope, etc. Then, in addition, charge a low hourly rate that covers the variable expenses. Don't forget to find out what the non-university differential is and mark-up the rate for off-campus customers. Billing is easier if you automate it by computer and its also easier to see how you did, so you can adjust your rate next year. Good Luck and keep good records.
At 07:47 AM 1/24/96 CST, Robin Griffin wrote: } } This recharge issue has recently come up here to. We are trying to comply } with the rules and are looking for ways to do it without hurting the lab. } My understanding is that } 1) we need to charge rates that only recover costs (no profit - not ever a } problem here) and } 2) we must charge all univerisity users the same rates } } I was considering taking the cost of the lab and charging grants a } percentage, sort of like an overhead that would cover the costs. Our } accounting department seemed to think this would be okay. I liked it } because I am worried that if we charge a fixed hourly rate, users would } minimize lab. use. Many of my expenses are fixed so less use only means } less bang for the buck. However, the U. of Hawaii writer seemed to think } this might be illegal. It is also very scary to hear that they will come } and audit- Eek! How about more specifics about the lump sum charging you } did that got you in trouble and your experiences with the audit. } } Another issue, we have some professors that support the lab when they have } money but some years they come up dry. We've always given them free time } during their dry years with the idea that they can't get more grants without } some work to show. It has always paid off for us. However, this appears to } be against the recharge center rules because we are charging people with } grants more than people without. } } One other issue, along with doing research, we teach laboratories on our em } lab instrumentation. Any comments on how or if this is billed? I can't } find anything in our rules about it. } } Thanks } } Robin Griffin } EM Lab Manager } Materials and Mechanical Engineering } University of Alabama at Birmingham } } Richard L. Markgraf Microscope Services University of California, Davis Ph. (916)752-3477 Fax (916)752-6363 rlmarkgraf-at-ucdavis.edu
Has anyone out there heard of a company called Burleigh Instruments Inc? If you have and they are still in existence we would appreciate it if you could let us know their address or phone number.
Thanks in advance.
Regards Richard.
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
I had some experience in thin sectionning in polymer materials for TEM.
The resin I used is water soluable GMA. The reference can be found on:
Zhiyu Wang etc., The study of microstructure and ultrafiltration membrane by electron microscope. Preceeding of the 1990 international congress on membrane and membrane prcess. Vol. II, pp. 1211-1213, Chicago.
Please let me know if you have question.
Zhiyu Wang Department of Biosystem Engineering University of Hawaii Honolulu Hawaii 96822
Original Subject: Burleigh Instruments Inc - address anyone?
Has anyone out there heard of a company called Burleigh Instruments Inc? If you have and they are still in existence we would appreciate it if you could let us know their address or phone number.
Thanks in advance.
Regards Richard.
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
******************************************************************************* *Tom Thielen "People who make sweeping generalizations are stupid!" * *Pre-Med Major * *Winthrop University thielent-at-lurch.winthrop.edu * *******************************************************************************
} At 9:13 AM 1/24/96, Griffin, Robin wrote: } } I'm trying to get in touch with TSL TexSEM Laboratories Incorporated a } } company that makes EBSP software for Windows. I have a phone number } } 801-467-9930, but nobody answers. Anyone know how to get in touch with } } them? } } } Didnt they get bought out by Noran? } } I heard a rumor..... } } John Mansfield
They weren't bought out by Noran, but rather Noran is marketing their systems. You can still deal directly with the TSL people if you want.
Bob Keller NIST Mat'ls. Reliability Div. Boulder, CO
} Esteemed Colleagues: } } I have been asked to image the interior structure of a polymer } suspension. }
**specifics deleted**
Any thoughts on the usefulness of freeze fracture to get a look at the internal structure? I'm not a polymer person either, so don't have any experience with this. You could, perhaps, avoid chemical changes with a freezing technique.
We are interested in setting up a Langmuir trough with fluorescence microscope. Currently, we are gathering information on fluorescence microscopes used for the studies of two-dimensional phase states of Langmuir monolayers at air/water interface. The microscopes are mounted on Langmuir trough and phase changes in lipid monolayers can be monitored with compression of the film. [e.g see reference K.J. Stine and C.M. Knobler, Ultramicroscopy 47, 23-34 (1992)]
Any information regarding types of fluorescence microscopes used in such studies, their resolution, particular requirements, and accessories, etc will be of great help. Thank you, Vitthal Vitthal Kulkarni, Ph.D. Membrane Biochemistry The Hormel Institute University of Minnesota 801, 16th Avenue NE Austin, MN 55912 Tel. 507-437-9626 Fax. 507-437-9606
J. A. Mccray D. R. Trentham Properties and uses of photoreactive caged compounds Annu. Rev. Biophys. Chem. 1989 18: 239 270
On Thu, 25 Jan 1996, William Tivol wrote:
} } I wonder what equipment is used or should be used for uncaging (photolysis) } } of caged (photoactivable) compounds. } } [snip] } } - What light sources are used (types of lasers, Xenon arc lamps, Mercury arc } } lamps, flash lamps)? } } } } - How is the exact light path to the cell/cells/subcells? What types of } } optics and components are used? } } [snip] } } Dear Martin and especially SPM'ers on this list, } Can this be done using either a NSOM probe or an NSOM-like input } probe? If so, this would likely have many uses. } Yours, } Bill Tivol }
I've gotten several off-line inquires about the Microscopy & Microanalysis - 1996 which is being held in Minneapolis on August 11-15,1996, and is the joint annual meeting sponsored by MSA, MAS, MSC-SMC . Many have asked about the fact that they have not yet received registration information.
I am posting this just to reassure all of you, that you will receive relevant the information, albeit slightly late this year. The registration booklet and call for papers is behind schedule this year and we anticipate it being shipped within 2 weeks. As soon as the booklets are ready for shipping I will post another announcement to let people know that they are "in-the-mail".
The information will also be put up on the MSA WWW site, probably by the middle of next week. However, you will not be able to download the various manuscript forms, since they will not be suitable for publication.
Abstracts of the Major Symposia and Topics for contributed papers/posters are already on that site (http://www.msa.microscopy.com) under the Annual Meetings Topic and you can use that information to start planning your manuscripts and which session you would like to submit them to.
The deadline for receipt of Abstracts remains March 15th as in previous years. Registration can be of course accepted right up to the day of the meeting, by snail-mail, fax, phone, or in-person.
Cheers... Nestor Your Friendly Neighborhood SysOp & Microscopy & Microanalysis - 1996 Program Chair
This is a good question to post to the microscopy listserver: Microscopy-at-MSA.Microscopy.Com (ask to have replies sent directly to you if you're not a subscriber). To subscribe, I think you send the subject & text "subscribe" to Listserver-at-MSA.Microscopy.Com (They've changed it since I subscribed.)
Richard Thrift p.s. again, I'm interested in the response you get. Thanks for providing the reference.
} } } Vitthal Kulkarni {vskulkar-at-wolf.co.net} 01/26/96 10:13am } } } We are interested in setting up a Langmuir trough with fluorescence microscope. Currently, we are gathering information on fluorescence microscopes used for the studies of two-dimensional phase states of Langmuir monolayers at air/water interface. The microscopes are mounted on Langmuir trough and phase changes in lipid monolayers can be monitored with compression of the film. [e.g see reference K.J. Stine and C.M. Knobler, Ultramicroscopy 47, 23-34 (1992)]
Any information regarding types of fluorescence microscopes used in such studies, their resolution, particular requirements, and accessories, etc will be of great help. Thank you, Vitthal Vitthal Kulkarni, Ph.D. Membrane Biochemistry The Hormel Institute University of Minnesota 801, 16th Avenue NE Austin, MN 55912 Tel. 507-437-9626 Fax. 507-437-9606
} Has anyone out there heard of a company called Burleigh Instruments Inc? If } you have and they are still in existence we would appreciate it if you } could let us know their address or phone number. } } Thanks in advance. } } Regards } Richard. } } } Richard Easingwood } South Campus Electron Microscope Unit } Otago Medical School } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } SOUTHERNMOST E.M UNIT IN THE WORLD
Try
Burleigh Instruments Inc Burleigh Park Fishers, NY 14453 (716) 924-9355 fone (716) 924-9072 fax 97-8379 tlx
UK Burleigh Inst Ltd Nine ALlied Business Centre Cold Harbor Lane Harpenden, Herts, AL5 4UT (0582) 766888 (0582) 767888 fax
Japan Techscience Ltd 0489 (64) 3111 0489 (65) 1500
Burleigh has written a very useful 17 page booklet that explains the basic of scanning probe microscopy called the Scanning Probe Microscope Book (part number (?) STM 364 493, 51993-0). I have found it to be a great intro to SPM.
Bob
Robert R. Wise, PhD Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
A technique we use to look at larger polymer particles that are sensitive to embedding in epoxies is to blend them with a film-forming latex, cast films and section.
-| Dr. John R. Reffner | email: rsrj2r-at-rohmhaas.com |- -| --------------------------------------------------------------|- The opinions expressed are those of the writer and not Rohm and Haas Company ________________________________________________________________
Esteemed Colleagues:
I have been asked to image the interior structure of a polymer suspension.
Ideally I would like to resin embed for TEM. I am not a polymer scientist, but I do know that these polymers produce spheres when suspended with small amounts of BSA. The polymer is polyglycolic acid (PGLA). I called the manufacturers, but they have not done any significant imaging of it, and do not appear to be interested. They did tell me that the PGLA is heat sensitive and dissolves in isopropal alcohol. My experiment with epoxys was an unmitigated disaster, and I am considering a low temperature embedment procedure (Another researcher tried Lowicryl, but the spheres did not remain intact).
I referenced a book entitled "Polymer Microscopy" by Sawyer and Grubb, but it doesn't refer to anything similar to my polymer.
Via: uk.ac.bbsrc; Fri, 26 Jan 1996 17:03:27 +0000 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Fri, 26 Jan 1996 17:05:10 +0000 X400-Received: by mta arcs.bbsrc.ac.uk in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Fri, 26 Jan 1996 17:05:10 +0000 X400-Received: by mta LARS.MUAS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Fri, 26 Jan 1996 17:03:05 +0000 X400-Received: by mta LARS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Fri, 26 Jan 1996 17:03:07 +0000 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; converted (ia5-text); Relayed; Fri, 26 Jan 1996 17:03:14 +0000
Dear Fellow Microscopists, Happy New Year to you all (not too late I hope!).
I routinely use a dilute solution of toluidine blue to quench some of the autofluorescence present in my material prepared for indirect immunofluorescence microscopy of tubulin. I am convinced that it does reduce the native fluorescence of my tissue (secondary vascular tissue of horse chestnut). However, I was floored at question time at the end of a talk at an international conference when asked, 'how does the quenching work?'. I have asked colleagues and consulted textbooks, but still do not have the answer. Can anybody help, please? [I'm sure that having the answer the question will never be asked again, but it might!]
Many thanks,
Nigel Chaffey: IACR - Long Ashton Research Station, Bristol BS18 9AF, UK [the Long Ashtonmost EM unit in the universe]
You can Low Viscosity Nitrocellulose from: Randolph Products Carlstadt, NJ 07072
Regards, Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
Nigel Chaffey wrote:} } I routinely use a dilute solution of toluidine blue to quench some of } the autofluorescence present in my material prepared for indirect } immunofluorescence microscopy of tubulin. I am convinced that it does reduce } the native fluorescence of my tissue (secondary vascular tissue of horse } chestnut). However, I was floored at question time at the end of a talk at an } international conference when asked, 'how does the quenching work?'. I have } asked colleagues and consulted textbooks, but still do not have the answer. } Can anybody help, please? [I'm sure that having the answer the question will } never be asked again, but it might!] } } Many thanks, } } Nigel Chaffey: IACR - Long Ashton Research Station, Bristol BS18 9AF, } UK [the Long Ashtonmost EM unit in the universe]
Brief rinsing with a very dilute solution of tol. blue also works to quench autofluorescence in Drosophila eyes, using thin frozen sections and FITC labelling. I'm not sure of the original rationale -the autofluorescence is very high, we were desperate for something that would work. I think just a hope that tol. blue probably binds to double bonds (??) and might just dampen things down..and so it did. Dont think I ever tried it with either rhodamine or a short-wavelength chromophore like DAPI ...does tol blue itself fluoresce under some excitation wavelengths?
Sally Stowe Australian National University EM Unit, Canberra. (28 degrees centigrade, blue skies, cool breeze..) ---------------------------------------------------------------------- Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 6 249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, |AUSTRALIA 0200
Some time ago I purchased a beautiful microscope. One of the lenses is an oil immersion type using "Cedar Oil". In a moments foolish enthusiasm, I immersed the lens in the oil and used it to my delight. Only after I was done did I realize that I have no idea how to clean the lens. Now the lens has a gummy oil residue left on it rendering it useless to me. Can anyone tell me how to clean my lens? I would be delighted to get an answer.
I recently obtained a B&L Model L photomicrographic camera with a bunch of lenses, base, bench, and accessories. It is equipped with a head which has a reflex viewer and accomodates 5"x7" film holders. I would like to be able to use this with 4"x5" film holders and thought perhaps someone on this list might have an old one around with some parts they'd be willing to part with.
Or, maybe someone can suggest where to get a reducing back to go from a 5"x7" to 4"x5" film holder.
If so, please drop me a note directly.
Thanks. Peter D. Barnett Forensic Science Associates - Richmond CA e-mail: pbarnett-at-crl.com FAX_510-222-8887
Message-Id: {199601290933.KAA23689-at-pons.uio.no} X-Sender: finnmog-at-pons.uio.no X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
know that they are new, more moderately priced, cooled, slow-scan cameras from Photometrics and Princeton. (The Photometrics SENSYS was announced last November and may only now be produced in volume - but it has been demonstrated in the US, I believe. The Princeton Micromax is a packaging of the Small cooled camera head and the ST133 controller).
If you have used or tested one or both for (biological) fluorescence microscopy, I should be grateful for the opportunity to put some practical questios, before deciding whether to go for one of them. (For the Princeton cameras, experience with the TE cameras and ST133/ST133 controllers is probably equally useful.)
Best regards, Finn-Mogens ***************************************************************** Finn-Mogens Haug University of Oslo, Institute of basic medical sciences, Department of anatomy E-mail: f.m.haug-at-basalmed.uio.no Box 1105 Blindern Phone : +47 22 85 12 67 N-0317 Oslo, NORWAY Fax : +47 22 85 12 78 *****************************************************************
} [snip] However, I was floored at question time at the end of a talk at an } international conference when asked, 'how does the quenching work?'. I have } asked colleagues and consulted textbooks, but still do not have the answer.
Dear Nigel, Fluorescence is the radiative decay of an excited state of a molecule to (usually) the ground state. Other decay modes exist and are always in competition with radiative decay--thus the fluorescence yield term in inten- sity formulas. Quenchers enhance non-radiative decay either by being accep- tors for energy transfer or by promoting collisional de-excitation, and by- and-large the energy transfer process is the most important. In this process, a nearby quencher molecule is converted into an excited state and the fluo- rescent molecule is converted into the ground state in a single step. The quencher then decays via a non-radiative process. Yours, Bill Tivol
The ISI (Topcon) DS-130 SEM at UC Berkeley has expired. Parts, i.e. electronics are needed. If you have or know of a DS-130 scope in storage or in a salvage state, please contact me. Thank you.
Doug Davis Staff Research Associate Electron Microscope Facility University of California Berkeley, CA 94720 (510) 642-2085 dbd1-at-uclink4.berkeley.edu
We always used to use a lens tissue moistened with a bit of xylene to clean the oil off oil immersion lenses, and this is the procedure recommended in an article, "Immersion oil and the microscope", by J. R. Cargille, President of Cargille Laboratories, the outfit that makes most immersion oils. In his article he points out that synthetic oils such as those manufactured by the Cargille Co. have a number of advantages over Cedar oil and other natural oils (i.e. they contain no volatiles, do not degrade from exposure to light and normal temperatures, etc.) If you'd like a copy of this article, send me your FAX number and I'll send it off to you. In any event, I think you should be particularly careful not to use acetone or alcohol, because they will soften and dissolve the cement that is usually used to hold the lenses in place. Best of all, check with the manufacturer of the lens.
Does anyone know of Macintosh or IBM PC software that is readily available (preferably free) for determination of lattice parameters from unknown x-ray powder patterns by the Rietveld method? W. C. Bigelow (bigelow-at-umich.edu)
} Esteemed Colleagues: } } I have been asked to image the interior structure of a polymer } suspension. } } Ideally I would like to resin embed for TEM. I am not a polymer } scientist, but I do know that these polymers produce spheres when } suspended with small amounts of BSA. The polymer is polyglycolic acid } (PGLA). I called the manufacturers, but they have not done any } significant imaging of it, and do not appear to be interested. They did } tell me that the PGLA is heat sensitive and dissolves in isopropal } alcohol. My experiment with epoxys was an unmitigated disaster, and I am } considering a low temperature embedment procedure (Another researcher } tried Lowicryl, but the spheres did not remain intact). } } I referenced a book entitled "Polymer Microscopy" by Sawyer and } Grubb, but it doesn't refer to anything similar to my polymer. } } Any (relevent) thoughts would be appriciated. } } Kathy Walters } CMRF } U of Iowa
Kathy,
Have you seen the article:"Cryo-Ultramicrotomy of Individual Latex Particles for Examination of Internal Morphology" by Angela M Marcelli? It was published in "The Microscope Vol 43:3 117-120 (1995)". If you supply your fax # I will fax you a copy.
L E Porter Phone (216) 796-1620 Head of Microscopy Fax (216) 796-3304 The Goodyear Tire & Rubber Company EMail LPORTER-at-GOODYEAR.COM Dept 415A 142 Goodyear Blvd Akron, OH 44305 USA
University of California Lawrence Berkeley National Laboratory Materials Sciences Division
National Center for Electron Microscopy Visiting Scientist Program
The National Center for Electron Microscopy is offering a fellowship that will allow participants the opportunity to conduct research in their own area of interest using the advanced transmission electron microscopes at the Center.
The program is intended primarily for young faculty/investigator electron microscopists, resident in the U.S. who are in the process of setting up their own facilities or are awaiting delivery of new equipment, and who could benefit from the head-start that use of instrumentation and interaction with personnel at NCEM would bring. However, other post- doctoral applicants with suitable experience and graduate students at an advanced stage of their thesis work would also be considered. Awards will be made according to the recommendations of the NCEM Steering Committee.
Fellowships will be of up to three-months duration and will carry a stipend of up to $6,000 to assist in defraying travel and living expenses. Applications must be received by March 31, 1996.
For further information and to receive application forms, contact
Gretchen Hermes, Coordinator National Center for Electron Microscopy, Bldg. 72 Lawrence Berkeley National Laboratory Berkeley, CA 94720 Tel.: (510) 486-5006 Fax: (510) 486-5888 Email: ghermes-at-lbl.gov
University of California Lawrence Berkeley National Laboratory Materials Sciences Division
National Center for Electron Microscopy Visiting Scientist Program
The National Center for Electron Microscopy is offering a fellowship that will allow participants the opportunity to conduct research in their own area of interest using the advanced transmission electron microscopes at the Center.
The program is intended primarily for young faculty/investigator electron microscopists, resident in the U.S. who are in the process of setting up their own facilities or are awaiting delivery of new equipment, and who could benefit from the head-start that use of instrumentation and interaction with personnel at NCEM would bring. However, other post- doctoral applicants with suitable experience and graduate students at an advanced stage of their thesis work would also be considered. Awards will be made according to the recommendations of the NCEM Steering Committee.
Fellowships will be of up to three-months duration and will carry a stipend of up to $6,000 to assist in defraying travel and living expenses. Applications must be received by March 31, 1996.
For further information and to receive application forms, contact
Gretchen Hermes, Coordinator National Center for Electron Microscopy, Bldg. 72 Lawrence Berkeley National Laboratory Berkeley, CA 94720 Tel.: (510) 486-5006 Fax: (510) 486-5888 Email: ghermes-at-lbl.gov
} [snip]... In any event, I think you } should be particularly careful not to use acetone or alcohol, because they } will soften and dissolve the cement that is usually used to hold the lenses } in place. Best of all, check with the manufacturer of the lens.
What is the reason for xylene being less likely to dissolve cement than acetone or ethanol? I would have thought that it depended on the cement and that generalizations would be difficult. Thanks for any insight on this.
We've used xylol for the last 30 years here with no serious consequences on any of our Zeiss and Leitz objectives. The service people use a combination of ether and other stuff, but we don't need it often enough to deal with having a can of ether on hand. Xylol should work fine, but never use alcohol or acetone on objectives. Mike ***************************************************************** Michael L. Boucher Sr. Boucher-at-TCRCA.USBM.GOV Geology-Mineralogy/Chemistry Labs Ph 612-725-4614 Twin Cities Research Center Fax 612-725-4527 U.S. Bureau of Mines Center 725-4500 Department of Interior 5629 Minnehaha Avenue South Minneapolis, MN 55417-3099 U.S.A. *****************************************************************
Does anyone know of database software that I could use to (1) input the form our lab uses for each sample that comes in (2) search the records by chosen field of information like "cell line" or "users name" for example and (3) build a new file with the records from that search? This would enable me to answer questions like "what were all the experiments done on x cell line from y time frame?" or place all the records from one user in one file that I could then print or give to him/her. I would also like to make a rapid index with just the identification number, date, user and cell or tissue type, as a rapid frame of reference.
Is there software to do this? Any recommendations or experiences?
Theresa
Dr. Theresa A. Fassel Sr. Research Associate fassel-at-post.its.mcw.edu Department of Microbiology (414)-456-8410 Medical College of Wisconsin Fax (414)-266-8522 8701 Watertown Plank Road Milwaukee, WI 53226-0509
I was wondering if anyone knows of any upcoming workshops in molecular biology using molecular probes for confocal and electron microscopy. I prefer a workshop where "hands on" techniques are emphasized.
Also, does anyone know where 22mm square, quartz coverslips can be purchased?
Dear all: I have attempted to respond directly to Alwyn Eades at 2 different email addresses listed in his 1-29 posting & get an error message of "illegal host/domain". So I apologize for posting this to the list.
I know someone who knows someone who bought a Philips SEM at our university's auction. I think he has given up on getting it set up. So he might be willing to sell parts from it. I don't remember the model no. It was one of only about 3 that were made with a motor-driven stage, about 1978 vintage. I have no idea any parts might be useable on your system. Contact me if you want to pursue it.
Nancy Smith nkrsmith-at-juno.com cc:smithn-at-uthscsa.edu 210-567-3861 Fax 210-567-3803
Hi Theresa - Regarding your inquiry, I would MOST strongly recommend "Q&A" for two reasons: 1) It is a true piece of cake to identify fields as "keyword" fields, then in each you can have an unlimited number of variables to search on - and build, if you want separate records, mailings, etc. The variables can be in any form. I happen to use codes (like "A") to indicate a particular category, but you can use anything - including full names. 2) Unlike the current highly regarded database systems (i.e. Foxpro, etc.) Q&A is a snap to learn. A half dozen hours of study, and you are up and operating. And, in addition, it is not very expensive. Good luck and regards, Don Grimes, Microscopy Today
Phil Rutledge writes "I was wondering if anyone knows of any upcoming workshops in molecular biology using molecular probes for confocal and electron microscopy. I prefer a workshop where "hands on" techniques are emphasized."
The 1996 EMBO course which is being held in Prague will have a session on *in situ* hybridization (as well as cryosectioning, immunolabeling and stereology). These courses are always "hands on" but the competition to get accepted on them is tough.
For more details contact: Dr. Ivan Raska Institute of Experimental Medicine AS CR Albertov 4 CZ-128 00 Praha 2 Czechoslovakia E-mail: raska-at-site.cas.cz Phone: +42-2-2491 0315 Fax: +42-2-294590
The original question referred to a situation where it was indeed necessary to clean cedarwood oil off an immersion lens. However, in routine use of an oil immersion lens, it has been my understanding over the years that it is best not to clean the lens. We use commercial non-drying immersion oil, and merely wipe the excess oil off the lens with a tissue after use.
A. Kent Christensen, University of Michigan, {akc-at-umich.edu}
-------------------------------
On Tue, 30 Jan 1996, Tobias Baskin wrote:
} Greetings, } Will Bigelow wrote: } } } [snip]... In any event, I think you } } should be particularly careful not to use acetone or alcohol, because they } } will soften and dissolve the cement that is usually used to hold the lenses } } in place. Best of all, check with the manufacturer of the lens. } } What is the reason for xylene being less likely to dissolve cement } than acetone or ethanol? I would have thought that it depended on the } cement and that generalizations would be difficult. Thanks for any insight } on this. } } Tobias Baskin } } - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - } ___ ____ ^ ____ _____ Tobias I. Baskin } / \ / / \ / \ / University of Missouri } / | / / \ / / Biological Sciences } /___ / /__ /_____\ / /__ 109 Tucker Hall } / / / \ ( / Columbia, MO 65211 USA } / / / \ \ / voice: 573-882-0173 } / /____ / \ \____/ /_____ fax: 573-882-0123 } } }
Back a few weeks to our discussion of flat bed scanners. It was noted that negatives need to be 1-2 stops overexposed for the transparency adapters to work properly.
UMAX has come out with a new version of there software that operates under Windows '95 that allows one to adjust the lamp intensity. I have not tried it. However one assumes that this would then enable one to compensate for light or dark materials that fall outside the range of the automatic settings -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
Scanning Microscopy International Post Office Box 66507, Chicago (A.M.F. O'Hare), IL 60666-0507, U.S.A. Telephone: (708) 529-6677 / FAX: (708) 980-6698 E.mail: 73211.647-at-compuserve.com
Scanning Microscopy, Cells and Materials, and Food Structure 1996 meetings will be from May 11 to 16 at the Hyatt Regency Hotel, Bethesda, MD (suburb of Washington, D.C.). Programs on the follow- ing topics are already being planned (*fliers available, list below): *Scanning Probe Microscopies (including STM, AFM, etc.) and Related Techniques for the Biological and Materials Sciences (program already has over 50 papers); Physical sciences programs in: *Fundamental Physics in Microscopy and Microanalysis (program of 20+ papers), *Pattern Formation and Nanoscaled Structures in Thin Film Formation (program of 25+ papers), *Scanning Microscopy and Semiconductors: Metrology and Diagnostics (Program of 25+ papers); Biological programs in: Microanalysis and Imaging, *Immunolabelling, *Radiation Effects, Apoptosis, *Dentistry, Corrosion Casting, *Inner Ear, *Bone Biology, *Stones and Crystals, etc.); *Cells and Materials (covering: Skeletal Tissue / Biomaterials; Foreign Body Reactions; Biointerfacial Reactions at Biomaterials Surfaces; Innovative Drug Delivery Systems; Blood Related Biomaterials; and Dental Biomaterials); and Food Structure related topics. For participation and contribution, please contact Om Johari at address above.
Scanning Microscopy International is also sponsoring another international meeting: 15th Pfefferkorn Conference on Electron Image and Signal Processing (immediately after the Bethesda Meeting) from May 18-22, 1996 at Silver Bay, New York (80 miles north of Albany, New York). The organizers are: Drs. Peter W. Hawkes (CNRS, Toulouse, France; FAX: 33-62-257999; E.mail: hawkes-at-cict.fr; as its chief organizer), W. Owen Saxton (Univ. Cambridge, U.K.) and Joachim Frank (NY State Dept. Health, Albany, NY). Nearly 45 invited contributions are planned; a flier is available on request. Interested contributors should contact one of the organizers.
I agree that xylene and alcohol are probably equal. An old Leitz Ortholux manual warned against using "spirits" to clean lenses. Modern cements are unlikely to be affected by either, but I still choose to use them sparingly. I always use 6" cotton-tipped applicator sticks and, after dipping them in xylene, press the bud against a tissue until it is just damp. It makes more sense than soaking the lens and cleans just as well. I also follow up with a dry bud when cleaning oil from a 40X objective. Then, I use the same method with Windex or other commercial glass cleaner to remove all traces and clean the lens thoroughly. Cotton tipped applicators have an enormous advantage over lens paper. They work better on small, concave, or recessed lenses, they aren't touched by your fingers, and so never transfer skin oils, and they prevent contact between your skin and any toxic solvents (such as xylene) you may use. I never use acetone on an objective. Some (mostly American Optical) have a painted "mask" around the lens, and acetone can dissolve the mask and deposit the paint onto the lens.
} Will Bigelow wrote: } } } [snip]... In any event, I think you } } should be particularly careful not to use acetone or alcohol, because they } } will soften and dissolve the cement that is usually used to hold the lenses } } in place. Best of all, check with the manufacturer of the lens. } } What is the reason for xylene being less likely to dissolve cement } than acetone or ethanol? I would have thought that it depended on the } cement and that generalizations would be difficult. Thanks for any insight } on this. } } Tobias Baskin } } - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - } ___ ____ ^ ____ _____ Tobias I. Baskin } / \ / / \ / \ / University of Missouri } / | / / \ / / Biological Sciences } /___ / /__ /_____\ / /__ 109 Tucker Hall } / / / \ ( / Columbia, MO 65211 USA } / / / \ \ / voice: 573-882-0173 } / /____ / \ \____/ /_____ fax: 573-882-0123 } } } } Richard L. Markgraf Microscope Services University of California, Davis Ph. (916)752-3477 Fax (916)752-6363 rlmarkgraf-at-ucdavis.edu
Mr-Received: by mta SPVX01.MUAS; Relayed; Tue, 30 Jan 1996 14:30:45 -0400 Mr-Received: by mta SPVX01; Relayed; Tue, 30 Jan 1996 14:30:46 -0400 Mr-Received: by mta SRVR01; Relayed; Tue, 30 Jan 1996 14:32:02 -0400 Disclose-Recipients: prohibited
Does anyone have any suggestions for placing fiducial points in tissues that are intended for embeddment, serial sectioning and immunolabeling? We are hoping to reconstruct serial sections as image stacks, ultimately for the purpose of measurement and visualization.
Some have suggested surface cuts, embeddment of threads, or even the inclusion of some kind of stainable implant.
} Does anyone know of database software that I could use to (1) input the } form our lab uses for each sample that comes in (2) search the records by } chosen field of information like "cell line" or "users name" for example } and (3) build a new file with the records from that search? This would } enable me to answer questions like "what were all the experiments done on } x cell line from y time frame?" or place all the records from one user } in one file that I could then print or give to him/her. I would also } like to make a rapid index with just the identification number, date, } user and cell or tissue type, as a rapid frame of reference. } } Is there software to do this? Any recommendations or experiences? } Dear Theresa, Any relational database program will do everything you ask. I have experience with dBASE, which works well, but is not free. All you have to do in a relational database program is to define fields with the requisite info, e.g., for the field "C_LINE" put in the cell line, etc., then use the search utilities to do boolian searches. The index file can be embedded in the main data file; you only need to print or list those fields in which you have an interest. dBASE can print these on forms which you can design to meet your own needs. As I said, almost any database program can do these things, so the one to choose is the one you can get most cheaply and/or which you can use best. Good luck. Yours, Bill Tivol
Subject: Time: 1:27 PM OFFICE MEMO More on Immers Oils Date: 1/30/96
Wow, did I open a can of worms when I mentioned the article on immersion oils by J. J.Cargille. Actually, the article was written in 1964, and is possibly quite a bit out of date by now. Basically it covers the following points: 1. describes the function of immersion oil in increasing the numerical aperture of a lens by increasing the refractive index between the objective and condenser. 2. Discusses the problems involved in formulating immersion oils, most of which are of no direct concern to users of them. 4. Notes that a variation of 1 C in temperature can change the refractive index of an oil by approx. 0.0004, so be sure to work at the temp given on the bottle 5. Synthetic oils can be formulated to have better properties than most natural oils; - freedom from color which may degrade performance - lower volatility and more resistant to oxidation and photo-decomposition, and so less likely to thicken and to form a gummy deposit on the lens. Wont thicken over time and change ref. index. - lower acidity and so less likely to damage your instrument, He also mentions that although Red Cedar oil has been one of the most commonly used oils: it has poor adsorption characteristics because it may be discolored; it contains volatiles and will leave gummy films on lenses unless cleaned off promptly, and it may thicken over time due to evaporation and change refractive index; it usually has a relatively high acidity, and so may be prone to damage the objective with prolonged use. He does not explain why it is common practice to use xylene to clean immersion oil off lenses, except to say that it doesn't damage the lens cement. I don't know whether this is the solvent recommended by all manufacturers of microscopes. In these times when such a wide variety of cements are available there may be some exceptions. However, I did check the instructions for a Nikon microscope we purchased only a couple of years ago, and xylene was recommended there. The rule I facetiously give to my students concerning the approach to using instruments is, "After all the controls are bent and everything is completely fouled up, read the instruction manual!" The present address of the Cargille Company is: R. P Cargille Laboratories, 55 Commerce Road, Cedar Grove, NJ, 07009 (Ph. 201-239-6633; Fx: 201-239-6096) if you want more up-to-date info on immersion oils. Sorry to get everyone so stirred up: Wil Bigelow (bigelow-at-umich.edu)
Subscribe Microscopy-at-MSA.Microscopy.Com MRG Associates, Inc. [Worldwide Headquarters] 755 Waverly Avenue Holtsville, New York, USA 11742 1-800-606-8869 (Voice within USA) 1-516-447-1041 (Voice) 1-516-447-1042 (FAX) E-mail "mrg1995-at-htp.net" O
Quite a few people have asked me for details of the federal audit at the University of Hawaii that deemed our billing practices fraud. Below is a brief summary that I sent to one interested person a couple of months ago--
} We apparently had two things going on here that annoyed the Feds. The } first, and the one that should be of the most concern to other } facilities that operate on a recharge basis, is that a Federal audit } alleged that we were making charges to Federal grants in advance of } receipt of services. We had been selling "shares" of instrument time, } allowing users to prepay for 80 hour blocks of time in advance. More } casual users who paid for time by the hour paid roughly twice as much } per hour as those who bought blocks of time. This was an incentive to } get people to commit to instrument usage, allowed us to collect monies } with which to pay our service contracts (which must be paid in advance), } expedited paperwork, and allowed users to encumber funds in a timely } manner. Everything went along fine for about 6 years, but then the Feds } decided that this constituted fraud. And they weren't very nice about } it, either. (As an aside, the auditors admitted that our bookkeeping } was exemplary and that the shares bit was a great and efficient idea and } should be allowed, but it isn't, so they slapped us anyway.) } } The second problem is related, and was actually the more serious on in } the Feds eyes, even though it involved less money. Apparently our } umbrella research unit transferred some funds to us as sort of a cross } between a retainer and a gift, to be used by one of the other research } labs if needed, but as a gift if not used. Unfortunately, they had us } write an invoice for it that said it was for use of the EMs, which were } never used. This was "clearly fraud". I guess if those monies had been } labelled as for core support or as a gift or whatever, it would have } been (almost) OK. The net result of these two practices was that about $20,000 was taken from us, presumably to be returned to the Federal grants } from which it came. Plus interest! In actuality, the $$ sits in some } account somewhere where no one knows what to do with it, and the matter } has been shelved. Meanwhile, we have allowed, over the last four years, } the users involved to continue to "work off" the money they had put into } the facility, at our loss. And so, with Federal and Hawaii state budget } cuts, we're in deep kim chee. The $$ that was taken from us would have } paid all our service contracts this year, for instance, and we are now } broke...! And we can't see breaking even for years to come. } I can see the Fed's points, they could see ours, and everybody lost on } this one. Don't let it happen to you!
Other things that must be considered include the federal granting agencies rules that you may not charge federal grants any more than the lowest rate you charge anyone else. With monies for any given project often coming from several sources, we quit trying to have differential charges altogether, so now we charge outside, commercial projects the same as internal U.H. projects. (Fortunately, there are no provate EM companies against whom we can't compete, so we don't have to worry about the politics of taking on any outside projects.) But then there could be a question about the taxpayers footing part of the bill for commercial users...
We had a site visit from a federal agency that supports roughly a third of our facility base costs (my salary included) two weeks ago. They seriously questioned why we didn't give preferential rates for users of the facility that were also funded by their agency. I'm confused, because if we have lower rates for their users, then other federal grantees would not have the lowest rates, and if we lowered them, then agency #1's rates would not be preferential, and so it would go until it was all free! BUT THEN one of the site visitors wanted to know what we would do if a faculty member had no money but needed to get preliminary data for a proposal, could we provide any services for free? And while one member was asking me that, another across the room was telling the facility manager that we CAN'T do any work for anyone for free because they were supporting us... They obviously don't have a unified plan.
The basic problem still remains that we have to pay our service contracts IN ADVANCE, but we have no reserve pool of money from which to do so (and some facilities are prohibited from accumulating a pool of money, anyway). And you can't collect money in advance of services rendered. Other recharge facilities here are in the same boat. It takes a lot of money for, say, DNA sequencing or protein synthesis, or whatever. The facility here would get the $$ up front to pay for gearing up to do it. That is now illegal. So they have to do it first (how?) and then charge the customer. And not make a profit to have the reserves to gear up for the next task...
Anyone have any words of wisdom?
Aloha nui, Tina } } ***************************************** } Tina (Weatherby) Carvalho * } Biological Electron Microscope Facility * } University of Hawaii * } (808) 956-6251 * } tina-at-ahi.pbrc.hawaii.edu * } http://www.pbrc.hawaii.edu/bemf/ * } ***************************************** p.s., Yesterday was gorgeous, about 83 degrees F, but today we expect rain, just to keep the mountains green.
Is anyone able to assist me in finding a CD ROM writer for image archieving in the price range of about 1000 $? Would be possibile to send me addresses where are those available?
Message-Id: {199601310905.DAA12836-at-Sparc5.Microscopy.Com} X-Sender: smgxstg-at-pop-server.bcc.ac.uk (Unverified) X-Mailer: Windows Eudora Version 1.4.4 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Going back a few years, as I do unfortunately, I remember the Microscope Service Engineer telling me never to use Xylene as it dissolved the mounting medium of the lens. He always used Ether.
The problem was he used to chuck the Ether soaked lens paper into the metal wastebin. He also used to smoke. And smoking wasn't banned in the Lab in those days. (I said I went back a few years). When he also threw his cigarette end in the bin, the result was exciting, (excessively so) to say the least.
It seems that there are quite a lot of different methods of cleaning lenses. My favourite method, for what it is worth, is to use either a proprietary mixture form Leica here in Sweden (isopropanol, some detergent and dist. water, I think) or abs. ethanol (both are OK from the manufacturer). After wiping the excess oil off around the lens (never on the lens), I put a drop of this on the lens, let it sit for a while, then blow it off in one go with clean compressed CO2. I repeat as many times as necessary. In this way I never need to touch the lens surface which according to many authorities (e.g. Shinya Inou=E9 in his book "Video Microscopy") may be detrimental as the surface coating of the lens is very soft and easily scratched or abrased even by "soft" cotton or lens cleaning paper. I prefer not to use solvents like xylene due to their potential health risks which I think should be considered in this context. Finally, it would be nice if reps. from each of the large microscope manufacturers could give the final words in this matter (what is best for the lenses, what can the cement take, what if there is other stuff than oil to clean (e.g. Moviol or other mountants), what is the best way to clean non-oil immersion lenses that have been messed up with oil or other things).
I have recently returned to Trent U and we have started to look at a computer archiving system which we would like to use in teaching. We have a small computer (MACS) lab which we are expanding. I am interested in finding out what systems other labs are using to grab images. Our aim is to grab images from an SEM, a CCD hooked up to a LM or Video camera and be able to enhance the images and add script and then download them through our LAND to the teaching lab. We also wish to do this all in colour. If you can give me any leads please reply.
We have been using Q-tip brand cotton swabs dipped in chloroform to clean our Zeiss objectives. The method works extremely well, and requires a minimum of scrubbing. Using this method over the past 7 years has resulted in no problems. I would caution that other cotton swabs may scratch the lenses. Doug Keene, Shriners Hospital Research Unit
I have a Fisons/Kevex 770 XRF system and would like to upgrade the computer systerm to a Windows based software system. Does anyone know of other sources of upgrade hardware/software other than the manufacturer? Thanks
I use Kodak lens cleaner on a cotton swab and blot with a clean cotton swab (Q-tip will do) for both oil immersion and non-oil immersion lenses. It's certainly of less of a health threat than xylene or chloroform and since it comes in a squeeze bottle is not likely to become contaminated by dipping cotton swabs into it.
Message-Id: {199601312216.QAA25413-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I am having a recurring problem with either my grid stain, or water in my knife boat, or wash water. We stain epoxy sections on copper grids with the usual uranyl acetate in 50% methanol followed by Reynold's lead citrate. These are washed in ultra pure water boiled to remove carbonate. We have done this routine stain procedure successfully for years and have not changed our materials or procedure in any way. However recently our sections have been plagued by almost perfectly hexagonal shaped deposits of electron lucent to opaque "stuff". These deposits aren't embedded within the epoxy as they ripple and burble under a concentrated beam. Microprobe (EDXEA) of the deposits gives a solitary lead peak (no uranium peak), but it is unclear whether the material is an inappropriate lead deposit or another material, that may have come from the boat or wash water, that is binding the lead stain. Between the hexagons, we see that the embedded tissue stains well. I would appreciate any suggestions as to what this might be and how to stop it.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
Dear All, Could you please, help me to locate E-mail and other contact items of Prof. William A. Miller, Professor of Oral Biology , some times ago at State University of New York at Buffalo. Please contact me directly on my Email, W.Jablonski-at-csl.utas.edu.au Regards to all, and happy New Year 1996, Wis Jablonski, OiC, EM/X-ray Microanalysis, CSL, Uni of Tasmania
I was recommended to clean any gunked up lens with a knob of polystyrene foam - no chemicals, just the dry foam rubbed in a circular motion onto the lens. Over the years none of my Zeiss lenses has suffered any damage from this.
Diana van Driel Dept Ophthalmology Sydney University 2006 NSW, AUSTRALIA
Considering the number of comments and questions I received concerning my recent recommendation of Q&A database software, kindly allow this response to the full list: The name of the software is "Q&A" and it is made by Symantec. Going back some 5-6 years, it was highly recommended by a number of the computer magazines. Pricing used to be listed by the software vendors in the back of these computer magazines, and I expect that they still do - or you might call Symantec direct (408)253-9600. To further explain its advantage, we all know that all database software has a common set of fields. And the normal application is to put only one variable in each field. For example, one might have a "last name" field - in which goes only last names. Theresa's question was to S/W that can conviently handle a considerable number of variables for each record. In Q&A during setup, you can designate a field as a keyword field. Then in that field you can have hundreds of variables - each separated by a semicolon. You can then search (include or eliminate) just as if your variables were in the dedicated fields. And you can have more than one keyword field - and search (include or eliminate) on variables from BOTH fields. For example, one of my keyword fields is "interests". I enter whether the person is a "user" or a manufacturer/supplier, his/her primary field(s) of microscopy interest (biology, material or earth science), and then his/her specific interests (electron, light, confocal, AFM, etc., etc.). In this one field I happen to have a total of 14 variables. If the person is a user I enter an "A", or if he/she is a manufacturer I enter a "B", if he/she is interested in electron and confocal microscopy, I enter an "C" and a "D", etc.- each code # followed by a semicolon. I could, of course, enter real text rather than the code. Say I want to find users only with an interest in biology and light microscopy. A piece of cake! And to extend the search to include only those who work in an individual company in a specific town is so, so easy. I do understand that other database software allows these kind of searches but all I have looked at, and I have looked at most, makes it somewhat to very difficult. I hope that a number of you find these comments of interest. Regards, Don Grimes, Microscopy Today
I would like to thank all of you who have responded to me personally with sympathy or ideas concerning our audit woes. Actually, we are OK right now (except for always operating in the red, which we are getting away with for now). My message was primarily to stimulate discussion on the subject of recharge problems, and as a warning to other facilities who may be subject to such scrutiny in the future. Rearranging our billing practices was not so awful; losing a chunk of money and being psychologically dragged over the coals was.
In my opinion, each facility that is not totally subsidized by their institution needs to take a good look at their sources of funds, and at what their outlay is. We basically have to justify every nickle of our hourly charges for instrument use, for example. We have built into that charge some percentage of the costs of consumables, technician salary for maintenance, cost of other down time, whatever. We do not add in a portion of the service contracts because they are (partially) supported by other sources. We should, however, add in that portion NOT covered by other sources. But the calculations have gotten so cumbersome as it is, that we have not. Plus there recently came up the question of use by faculty partially supported by the same grant as supports part of the service contract, and should that be prorated? It gets unbelievably complex really quickly. So I'm just thorwing this out there to (make your lives more miserable, and) warn others of the potential budgetary/political problems that may come out of the blue. I expect each situation to be different. I am interested in finding ways to go back to the "lump sum" situation for our regular users. For instance, perhaps they can pay outright for a percentage of the service contracts, and then not be charged by the hour. It probably all depends on the current person sitting in the fiscal or contract office, or what they are wearing, or whatever. I'm interested in what works for other facilities.
Our rainstorm blew through, and today it's 83 degrees, clear and sunny. And I'm stuck in this EM lab...
***************************************** Tina (Weatherby) Carvalho * Biological Electron Microscope Facility * University of Hawaii * (808) 956-6251 * tina-at-ahi.pbrc.hawaii.edu * http://www.pbrc.hawaii.edu/bemf/ * *****************************************
Thanks, Tina, for sharing your woes with the readers of the microscopy list.= As was discussed at the Technologist's Forum at MSA last year, these= issues affect all EM service labs that deal with federally funded research= projects. The answers are not always easy, but there are some basic= guidelines that we use at Colorado State University that could be applied= at other universities.
** some deleted **
} In my opinion, each facility that is not totally subsidized by their=20 } institution needs to take a good look at their sources of funds, and at=20 } what their outlay is. We basically have to justify every nickle of our=20
This is what we have found. Financial support of EM labs from central= sources, like college deans and vice presidents for research is essential. = When we polled about 2 dozen EM labs at universities comparable to ours a= few years ago, the clear message was that to not have central support was= "the kiss of death".
} hourly charges for instrument use, for example. We have built into that=20 } charge some percentage of the costs of consumables, technician salary=20 } for maintenance, cost of other down time, whatever. We do not add in a=20
We have had to do rigorous cost accounting to find out as accurately as= possible what the hourly cost of operating the instruments is. We know= what the fixed costs for each instrument are. They are service contracts,= gases, liquid nitrogen, an estimated number of hours for routine= maintenance, etc. Those expenses are divided by the expected usage, in= hours, to come up with an estimated hourly charge. Our best estimate for= our instruments is $45/hr in actual cost to run the instruments.
We have gone through several mechanisms over the last 5-10 years to get= financial support from our central administration. What happens this year= and next is that several central sources have paid enough dollars to cover= some of the fixed costs, specifically service contracts, which allows us to= bill every user at CSU the same reduced rate of $17/hr. There are three= important points to remember; 1) no dollars from federal grants are used= for the up front payments, so there is no pre-payment for services, 2) all= CSU users are billed the same amount for the entire fiscal year, and 3) the= hourly charge reflects real expenses.
This entire process is a collosal pain, but it is essential, if we want to= stay in compliance with federal regulations. We think the effort will= serve us well, should we be audited. We have been working closely with the= financial officer of our college and the university's controller to be as= sure as is possible that our policies will pass their scrutiny, as well as= that of auditors.
These policies aren't used only in the EM Center, but also at other centers= around the university, and will be used by more. So what happens at other= labs at your university might be useful to you.
} portion of the service contracts because they are (partially) supported=20 } by other sources. We should, however, add in that portion NOT covered=20 } by other sources. But the calculations have gotten so cumbersome as it=20 } is, that we have not. Plus there recently came up the question of use=20 } by faculty partially supported by the same grant as supports part of the=20 } service contract, and should that be prorated? It gets unbelievably=20
A multi-tiered fee schedule invites such headaches that we won't even= consider it. The bookkeeping is so cumbersome and convoluted that we think= it invites trouble.
Our philosophy is that we will bill for real expenses fairly, so no one gets= gouged. We had been subsidizing the campus for a long time, and, once we= decided to clean up the books, we made everyone acccountable.
We are trying to meet our expenses as closely as possible, without making= money. If we end the year with a deficit, we can legitimately charge that= against next year's budget and recoup it through adjusted fees next fiscal = year.
} complex really quickly. So I'm just thorwing this out there to (make=20 } your lives more miserable, and) warn others of the potential=20 } budgetary/political problems that may come out of the blue. I expect=20 } each situation to be different. I am interested in finding ways to go=20 } back to the "lump sum" situation for our regular users. For instance,=20 } perhaps they can pay outright for a percentage of the service=20 } contracts, and then not be charged by the hour. It probably all depends on= =20
This is the kind of arrangement that we think is sure to get us in trouble,= so we won't consider it. Once the true costs of running the facility are= known by the users, they are much more likely to go along with the charges.= Everyone would like to get the services for free, but it's hard to argue= with facts like real costs.
So far, this approach is working very well. And, since we're still looking= for better ways to recoup expenses, I'm interested in any other procedures= that work.
} the current person sitting in the fiscal or contract office, or what they= =20 } are wearing, or whatever. I'm interested in what works for other=20 } facilities.
} I have a Fisons/Kevex 770 XRF system and would like to upgrade the computer } systerm to a Windows based software system. Does anyone know of other } sources of upgrade hardware/software other than the manufacturer? } Thanks
Dear Don, The company IXRF have contacted old Kevex EDS users with a system to run EDS software on a Pentium computer, using the power supplies and pulse processor of your existing Kevex. With a name like IXFR, I suspect they do XRF systems, too. Contact: Kenny Witherspoon IXRF Systems, Inc. 15715 Brookford Drive Houston, TX 7705 tel: 713-286-6485, fax: 713-286-2660
They will probably be at MAS/MSA/MSC, too. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
} Does anyone know of database software that I could use to (1) input the } form our lab uses for each sample that comes in (2) search the records by } chosen field of information like "cell line" or "users name" for example } and (3) build a new file with the records from that search? Dear Theresa, I have been using Microsoft Access for keeping track of my billing in my lab for about three years now. It is a relational database and you can design the particular "queries" to relate any data to any other. I do Christmas cards at home, keep track of SEM hours, photos, different customers and rates at work. I know it will do a lot more than I use, and I find it easy to change things if I need to.
I suspect any good database would be sufficient for your use, it is just a matter of choosing one. The learning curve is a bit slow at first, but you soon learn.
Hope this helps, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Message-Id: {199602011357.JAA16914-at-Snoopy.UCIS.Dal.Ca} Comments: Authenticated sender is {RMACKAY-at-ac.dal.ca}
To those in probe land,
Occasionally I am asked to identify an unknown mineral based on microprobe data. Does any one know of an inexpensive software program that would run on a PC ?
Best Regards,
Bob MacKay Robert MacKay Department of Earth Sciences Dalhousie University Halifax, Nova Scotia, Canada B3H 3J5 Tel: 902 494-7087 e-mail rmackay-at-ac.dal.ca
I have been thinking about a similar problem with respect to serial sectioning of the hippocampus. Could the use of a microbeam laser to produce a micron-size hole in the tissue be part of a solution? Leo Marin Dept. of Physiology University of Toronto
Message-Id: {199602011604.KAA15966-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 08:43 AM 2/1/96 -0500, you wrote: ......My conclusion was that the Pb was present as an impurity on the grids, some batches have it, some don't. } ************* That's something I hadn't thought about sinse we routinely wash our grids (copper mesh) in acetone followed by alcohol. We will try an acetic acid wash next followed by thorough rinsing in clean water and see what happens.
Message-Id: {199602011604.KAA15957-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 08:37 AM 2/1/96 -0500, you wrote: .....perhaps the problem appeared after the resin cartridges were exchanged and hence } most likely to leach chelating agent. } *************
That's a good point. However the water used is from a reverse osmosis system and shouldn't have that problem. Also up until the recent past, we had indeed used resin demineralized water without problem. Thanks for your input.
Message-Id: {199602011604.KAA15980-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 09:20 AM 2/1/96 EST, you wrote:
.....we thought it was coming from the lead solder in the water pipes.....you might ask Don Steel at alcan.....
************** Paul -
I just read Don's letter and am going to try his suggestion to wash grids better. We have had this problem with both resin demineralized water as well as water purified by reverse osmosis that is used in tissue culture. The success of our tissue cultures seems to preclude a significant lead content in the latter.
I must admit that I feel a bit better hearing that others have come up against these -at-#%&*-at-#$! hexagons. * * Joiner Cartwright, Jr. * *
Message-Id: {199602012017.OAA21191-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 10:10 AM 2/1/96 -0800, you wrote:
} The hexangonal crystals are probably uranyl acetate, not the lead. Take } a look at your Uranyl staining procedure. The stain is probably over } saturated or drying out at some point or possibly too old. } } Dan
********************** Dan -
I'll check that out and see what happens. However EDXEA microprobe of the contamination yielded a lead peak only.....but, shoot, I'll try anything at this point.
In response to Joiner Cartwright's question about a precipitate that is forming on his grids when he uses "ultra pure water boiled to remove carbonate", I have the following suggestion:
I was taught to never use resin-purified water in solutions for EM work. The rational was that chelating agents in the resin may become solubilized and later create unwanted precipitate in the specimen. I only use glass distilled water for all of my EM work. Perhaps the problem suggested above is due to the water (I assume that "ultra pure" refers to a resin purification system. If this is the case, perhaps the problem appeared after the resin cartridges were exchanged (and hence most likely to leach chelating agent).
__________________________________________________________________________ Donald L. Lovett e-mail: lovett-at-trenton.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 Trenton State College, NJ 08650-4700 fax: (609) 771-2674
} Could the use of a microbeam laser to } produce a micron-size hole in the tissue be part of a solution?
Dear Leo & all, Since the object is to produce contrast between the rest of the section and the fiducial mark, a micron-sized hole would do the job. One must remember that during the fixing, staining & embedding the hole might be bent or otherwise distorted, but that information is useful in itself. An alternative to using a laser is to use a micropipette. We have been developing micropipettes for HVEM sample holders & have discovered that they can be pulled to submicron diameters, so they may offer an advantage over lasers. (Good old low-tech solutions still have merit. :-)) Yours, Bill Tivol
Message-Id: {199602012117.PAA28325-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Thanks, Chuck. That sounds plausable. We'll give it a try.
Joiner ************************************
At 02:52 PM 2/1/96 -0600, you wrote: } Joiner, } I posted a message to Microscopy and never saw it come over the } newsgroup. So anyway.... } We have had the same problem. If we use fresh sodium citrate in } the preparation of the lead citrate, it goes away. Information, from ? } somewhere, indicated that sodium citrate can degrade over time. It isn't } easy buying small quantities of the stuff so we buy 500 grams and aliquot } out the powder in 15 - to 20 gram portions in scintillation vials and keep } it out of the light. When we use a vial, we discard any leftover. If two } vials/two batches of Pbcitrate show problems, we discard any remaining } vials and order a new batch of sodium citrate. } We may be fooling ourselves and the leftover sodium citrate is an } adequate sacrifice to appease the quirky gods of EM. Either way, it works } for us. } } Chuck } } } } Charles J. Butterick (Chuck) } Electron Microscopy Center } Department of Cell Biology } and Biochemistry } Texas Tech University Health } Sciences Center } 3601 4th Street } Lubbock, Texas 79430 } } vox (806) 743-1633 } fax (806) 743-1219 } email emccjb-at-ttuhsc.edu or } chuck-at-micron1.lubb.ttuhsc.edu } } } }
Joiner Cartwright said, "I am having a recurring problem with either my grid stain, or water in my knife boat, or wash water. We stain epoxy sections on copper grids with the usual uranyl acetate in 50% methanol followed by Reynold's lead citrate. These are washed in ultra pure water boiled to remove carbonate. We have done this routine stain procedure successfully for years and have not changed our materials or procedure in any way. However recently our sections have been plagued by almost perfectly hexagonal shaped deposits of electron lucent to opaque "stuff". These deposits aren't embedded within the epoxy as they ripple and burble under a concentrated beam. Microprobe (EDXEA) of the deposits gives a solitary lead peak (no uranium peak), but it is unclear whether the material is an inappropriate lead deposit or another material, that may have come from the boat or wash water, that is binding the lead stain. Between the hexagons, we see that the embedded tissue stains well. I would appreciate any suggestions as to what this might be and how to stop it."
I've had the same problem. Generally the problem occurs when our sodium citrate gets too old. When we order fresh sodium citrate, we aliquot 15-20g into separate scintillation vials and seal the vials tightly. We use the amount necessary to make Reynold's lead citrate and dispose of the leftover sodium citrate in the used vial. Apparently, the reopening of a bulk container allows degradation of the sodium citrate. Then again, this might be silly superstition and my gift of leftover sodium citrate might be a suitable sacrifice to appease the fickle nature of the gods of EM.
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
Fourth Annual Microscopy Colloquium hosted by California State University and Southern California Electron Microscope Society
Saturday, May 4, 1996 Sunday, May 5, 1996
at the San Diego State University Aztec Center, San Diego California
Business meeting for CSU delegates Friday, May 3, 1994
Abstracts Due March 15, 1996
The proceedings of the meeting will be published in Microscopy Research and Technique.
This Colloquium provides a forum for the exchange of research interests and experiences in all fields of light and electron microscopy, in biological, geological, and materials sciences. Participants include students and scientists from academia and industry. Both platform and poster presentations are invited.
Student presentations are strongly encouraged
Registration:
Registration Fees: $35 Regular $10 Student $50 Vendor by March 15
Late Registration: $45 Regular $10 Student $60 Vendor, after March 15
Makes checks payable to: EM Facility/Colloquium
Send to: Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego, CA 92182-4614
For additional information, flyers, and abstract forms, contact: Steve Barlow (619) 594-4523 or sbarlow-at-sunstroke.sdsu.edu
--------------------------------------------------------------------------- ------------------------------------------------------------------------------ Dr. Steve Barlow EM Facility/Biology Dept. San Diego State University San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu
Dear All, Thank you very much for helping me to locate Bill Miller and especially Prof. Summers, Irving Delilah and Susan Udin.Thanks again. Cheers, Wis Jablonski , CSL, Uni-Tas, Australia
I was advised by my microscope rep. (a very respected microscopist) to use a heavy polishing compound (like used to remove paint on cars) and light grit sandpaper to remove oil and lens coatings on our lenses. On particularly difficult occasions a sandblaster has sufficed. Needless to say that in 23 years we have experienced no difficulty using this method. We have never published a paper based on our findings but we hope to get a Science article soon.
I thought that with the huge influx of postings regarding this issue I would just add my 2 cents worth as it seems everybody had a different idea about how to do it. You should contact you local microscope rep. if you need to know how to clean a lens.
Altera Corporation SEM/EDX Technologist (Reliability Technician Opportunity)
COMPANY: Altera Corporation is a leader in advanced programmable logic devices and software. Our continued growth and success necessitates the need for the following talented individual to join our team.
JOB DESCRIPTION: In this position you will work with product, reliability and technology engineers to identify appropriate failure analysis techniques, provide physical analysis service to identify the root causes of the failures,
document the results, and inform the engineers of the root causes for corrective actions. You will also coordinate the maintenance of failure analysis equipment.
QUALIFICATIONS: The ideal candidate will have one plus years related experience in physical analysis. Also they will be familiar with usage of analysis equipment including SEM, EDX, precision cross sectioning, RIE, Jet Etcher and chemical de-processing set-up. Good communication skills, ability to handle multiple projects and interact effectively with others is a must.
BENEFITS: Altera offers a challenging environment along with comprehensive benefits which include profit sharing, company matching 401(k) plan and an Employee Stock Purchase Plan.
CONTACT: Principals only please. EOE. Send resume to:
Altera Corporation, Human Resources, Attn: JG/SJDC295, 2610 Orchard Parkway, San Jose, CA 95134-2020. FAX: 408-435-5065.
} Diana van Driel wrote: } I was recommended to clean any gunked up lens with a knob of polystyrene } foam - no chemicals, just the dry foam rubbed in a circular motion onto the } lens. Over the years none of my Zeiss lenses has suffered any damage from } this.
Have you inspected the front lenses of your objectives under magnification to see if there are scratches in the coating? If there are none, is is a testament to the durability of the Zeiss coatings.
I have seen too many microscope lenses (yes, even Zeiss) marred by rubbing with dry materiels. I don't recommend this for any lens. The dust particles that settle on lenses, particularly uncovered oculars, act like sandpaper when rubbed across the lens surface. Solutions such as lens cleaners lubricate these particles and reduce their abrasive effect. Cotton buds tend to lift the particles into their fibers, while, I believe, polystyrene would tend to keep them at the lens surface where they can do damage.
This discussion of cleaning gunked immersion lenses misses the real problem, in my mind. I care for 2100 microscopes constantly in use by university students. We use only synthetic immersion oils. Despite warnings, a certain number of these microscopes end up with immersion oil on the high dry (40X) objectives, rendering them useless until cleaned. Since the microscopes may be of 8 different brands, of varied models and ages, my cleaning method has to work for them all, and be applicable in a classroom situation. Cotton buds and glass cleaner (Windex or something similar) used in a wash and dry pattern, work well for cleaning all lenses and is relatively benign to microscope and student. Glass cleaner doesn't work well for immersion oil on the 40X, because it requires repeated application of the glass cleaner to remove all traces, a time-consuming effort. The US Government, and particularly the state of California, have stringent safety rules governing use of chemicals. While xylene, may be the best cleaner for immersion oil, its toxicity is a drawback and I hesitate to recommend it for students in a classroom environment. Same goes for pet ether, methyl-ethyl ketone, acetone or chloroform. Alcohol would be a preferred solution, but, because of warnings about its use with older microscopes, I hesitate to recommend it. Perhaps when the older microscopes are completely phased out of our inventory ... Immersion oil seems to be a popular thread with this list. I would appreciate any comments or ideas.
Rick Markgraf Microscope Services University of California, Davis rlmarkgraf-at-ucdavis.edu PH. (916)752-3477 FAX (916)752-6363
There is an immediate opening in our group at Northwestern University for a Postdoctoral Scholar or a Research Associate, in the area of Analytical Electron Microscopy/Materials Science. I would greatly appreciate it if you could pass on the following information to interested parties. Please have anyone interested contact me at the address/e-mail/phone below.
POSITION OPEN: Postdoctoral Scholar or Research Associate
Area: Analytical Electron Microscopy & Materials Science
Qualification: A PhD in Materials Science/Physics or related area. Very strong hands-on experience in various AEM techniques is required, including transmission EELS, x-ray microanalysis and CBED. Experience in specimen preparation of variety of materials, especially x-sectional TEM is required. Preference will be given to those who have demonstrated skills and publications using FEG TEM/STEM/SEM and interfacial/defect phenomena in solids.
Instrumentation available at NU in the newly restructured Electron Probe Instrumentation Center (EPIC) includes: A Hitachi HF-2000 FEG TEM/STEM with x-ray, EELS, in-situ IV/LHe holder and e- holography set-up, a Hitachi S4500-II FEG SEM with x-ray, EBSP/OIM and LHe stage with in-situ IV probes, a Hitachi H8100 cTEM, and access to various other TEMs/SEMs and other analytical instrumentation.
Research involves use of diverse AEM techniques to probe problems of thin film growth, interfaces & grain boundaries in oxide systems.
The position is open immediately for at least one year, renewable upon mutual agreement for longer period. Salary and benefits will commensurate with experience and skills.
Please contact immediately:
******************************************************* (Vinayak P. Dravid) Associate Professor, Materials Science & Engineering Director, Electron Probe Instrumentation Center (EPIC) Northwestern University, Evanston, IL 60208, USA Ph.: (847) 467-1363, Fax: (847) 491-7820 E-mail: v-dravid-at-nwu.edu *******************************************************
Thanks,
(NOTE THE NEW AREA CODE) ******************************************************* (Vinayak P. Dravid) Associate Professor, Materials Science & Engineering Director, Electron Probe Instrumentation Center (EPIC) Northwestern University, Evanston, IL 60208, USA Ph.: (847) 467-1363, Fax: (847) 491-7820 E-mail: v-dravid-at-nwu.edu *******************************************************
I need help in either getting articles or article references on a good, short EM paper that would be useful for teaching students. In the July-August 1992 American Scientist Magazine, there was an excellent article, entitled "The New Vision of Light Microscopy". This is an excellent paper on LM and image analysis for students and also for technician references. If anyone out there has any suggestions or ideas, please let me know. Thank you in advance. Cathy Kelloes
In the past I have used chloroform routinely for expanding thin sections. Because of the hazardous nature of this chemical I tried a heat pen (amps - .125), but it was totally ineffective. Would a more powerful heat pen be effective in reducing compression ? I would appreciate some feed back on this. Thanks Leo
While discussion is going on about the precipitate that is forming on grids even when ultrapure boiled water is used (Cartwright, Lovett). I have another question. I see sometimes grids jumping in plastic dish becose of static electricity. I imagine that if we place charged grid on stain solution that some stain will precipitate.Is out there anybody who would comment for this possibility? John Gabrovsek
Message-Id: {199602021559.JAA21589-at-BCM.TMC.EDU} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 04:28 PM 2/1/96 -0600, you wrote:
"Do you wet your grids before putting them in your methanolic uranyl acetate?"
No, we haven't been wetting the grids first. But that sounds like a reasonable thing to do. If Mollenhauer suggested it, it must be good.
*********************
"I also have one caution about using acetic acid to clean grids. If you leave the grids in the acetic acid too long, you'll get flakes of copper on your sections."
Yes, we're careful about that. One thing that may be contributing to the problem is that we're using the thin bar "Super 200" grids because we need to maximize the open space. These grids are difficult to wash as thoroughly as we'd like because there is less metal for the sections to adhere to and they are easily washed off the grid. But we've been using these grids and stain procedure successfully for years. We are looking for things in the laboratory environment that have recently changed. Our pay scale is not high on the list of suspects.
Greetings all! I am trying to use freeze substitution to retain soluble oligosaccharides in plant tissues, and am not yet convinced that I have been successful, partly because of embedding/infiltration problems (I think!). We have tried a variation of the tannic acid fixation with Kaeser's substitution cocktail, and stuff seems to look ok (tough to really assess when the scope has been down, unfortunately), although we have been advised that probably LR White is easier to work with than Lowycryl K4M, which is what we used the last time, and it doesn't section happily. Would like to try another run (just got some fresh LR White) and thought it would be useful to ask for some input from the group. It's probably of interest to more than me, but if folks prefer to respond directly, I will summarize the responses and post them. I'm certainly looking forward to your input!
TIA shea
S.Shea Miller Agriculture and Agri-Food Canada Centre for Food and Animal Research Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: 613-759-1760 Fax: 613-759-1765 email: millers-at-em.agr.ca
Message-Id: {199602021914.NAA17379-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 08:55 AM 2/2/96 -0800, you wrote:
} Could the UA somehow have been cross-contaminated with Pb? } Sometimes a weak wash with 0.02 N NaOH will help with Pb deposits-if that } is the source of the problem. } } } Doug Davis } ********************** Doug,
We've been pretty careful not to cross contaminate. We may have to resort to after-the-fact cleaning, but I would like to eleminate the problem before it occurs. Thanks for the suggestion.
I think there is something about the circumstances described over the last few days by Tina Carvalho that recalls some of the recent discussion of ethical values.
No matter how high and noble our calling, we all work within rules. Some of them are institutional, some professional and some come from the laws of our state and our nation. Whatever their source, however, the rules are what the rules are.
If we don't like the rules, we can work to change them, but none of us has a license to violate them. Auditors, internal or external, are in the business of finding violations of the rules, and some of them do a very good job of doing just that. All of us are tempted from time to time to bend or to break the rules under which we operate, sometimes for a very good purpose. In the case of our microscopy supplies business, for example, we may be asked to sell the components of a sputter coating system separately so that they can be bought out of a budget intended for consumables, when the rules say that the system is capital equipment, to come out of another budget. Is this right? Or is it a conspiracy to defraud? Does the noble purpose (the fact that the funds will go back to the sponsor) make any difference? How about the request that we predate or postdate the invoice, to get it into the "right" year?
It seems to me that we are all better off if we just force ourselves to live within the rules instead of trying to rationalize our efforts to get around them.
BTW, am I the only person who wonders which is the chicken and which is the egg when Tina comments about the lack of commercial EM facilities within her state?
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. 63 Unquowa Road Fairfield, CT 06430-5015 Ph: 1 203 254 0000 FAX: 1 203 254 2262 e-mail: AWBlackwoo-at-aol.com WWW--http://mail.cccbi.chester.pa.us/spi/spihome.html
Does anyone know where I might be able to purchase some used double tilt top entry specimen holders for a JEOL 4000 EX? Is there someone decommissioning a JEOL 4000EX that would like to sell their specimen holders?
Roseann Csencsits Argonne National Laboratory Argonne, IL 708-252-4977
Does anyone have a protocol for using (HPMA?) instead of propylene oxide and embedding coverslips directly in cell wells, and then using liquid nitrogen to remove the coverslip after plastic is polymerized?
Have you considered fixing the tissue in aldehyde and embedding in one of the other (hydrophobic) Lowicryl resins (e.g. HM 20, HM 23). They polymerize at lower temperatures and cut very well. We have had some success with soluble proteins in the pancreas and there are published examples of this method.
Here are a few references which might be of interest: van Genderen et al, 1991 J. Cell Biol. 115:1009-1019. Oprins et al, 1994 J. Histochem. Cytochem. 42:497-503.
There is also a recent paper in Microscopy Research & Technique which covers this subject but omits to refer to the above papers. The paper is in MRT, 1996 33:251-261. Look too at the work by Humbel & Schwarz which is cited in the MRT paper.
} 2 February 1996 } } From: Andy Blackwood } } To: Microscopy BB } } I think there is something about the circumstances described over the last } few days by Tina Carvalho that recalls some of the recent discussion of } ethical values. } } No matter how high and noble our calling, we all work within rules. Some of } them are institutional, some professional and some come from the laws of our } state and our nation. Whatever their source, however, the rules are what the } rules are. } } If we don't like the rules, we can work to change them, but none of us has a } license to violate them.
On the contrary. The idea that the existence of arbitrary rules defines ethical behavior is incorrect by construction. The classic examples, of course are in the realm of human rights violations -- where the ethical actions are explicitly those which are against the rules imposed by an unethical or irrational government.
It is *not* necesarily unethical to break rules, particularly when those rules are arbitrary, capricious, or in themselves unethical.
} It seems to me that we are all better off if we just force ourselves to live } within the rules instead of trying to rationalize our efforts to get around } them. }
When you call efforts to get around irresponsible regulations "rationalization" you beg the question that sometimes getting around the regulation is the only ethical thing to do. Sometimes finding an excuse for observing the regulations, and thus avoiding actions which involve personal risk, is the rationalization.
Ethics and rules are not orthogonal, but I, at least, refuse to let arbitrary bureacrats and supercilious functionaries get away with calling me "unethical" if I am not sufficiently obsequious to their dictates.
} Considering the number of comments and questions I received concerning my } recent recommendation of Q&A database software... } Don Grimes, Microscopy Today
I have used Q&A, and must agree with Don Grimes. Most database programs have a lot of unnecessary bells and whistles.
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
On February 20 was written:
} Subject: Pt TEM grids } } Does anyone make Pt TEM grids? They are needed for high temperature } experiments-other grids oxidize. } } Roseann Csencsits } Argonne National Laboratory } Argonne, IL } 708-252-4977 } } Pt grids are not in our printed catalog but are to be found in our all- electronic catalog on our WWW website as indicated below, in several different mesh sizes.
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: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
I am in the proces of purchasing a digital image acquisition system for a Hitachi H7000 stem. To do this I need a digital beam control box to interface the microscope to the acquisition system. The later models, eg. Hitachi H7100 have this circuitry built in but the older models (H7000) need the DBC box which Hitachi made but no longer does. Does anyone out there have one that they don't use anymore or know of any extras collecting dust on a shelf somewhere? I am slightly desparate and have some money (not much) to pay for such a beast. Thanks in advance for info or sources, Hank Adams EML, New Mexico State Univ email: hadams-at-nmsu.edu phone: 505/6463600
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. wrote: "......It seems to me that we are all better off if we just force ourselves to live within the rules instead of trying to rationalize our efforts to get around them........".
Andy is correct. The rules and laws must be followed. Those we don't agree with we can work to change. But am I the only one who reads Andy's entire commentary (that only mentions what academics do to get around some regulation in order to purchase an item) and feels resentment? I have been approached many times by vendors who ask me to write lock-out specifications so I will buy from only one source. Other vendors have informed me that if I bought a certain dollar amount of supplies, they would throw in the instrument for free that uses those supplies. More than once I have successfully appealed to the State of Texas on a bid where an unscrupulous vendor misrepresents their instrumentation and/or the price. Some vendors are as eager to earn the academic dollar as the academic is to stretch what little he/she has. What about vendors/business people who appeal to the government and/or granting agencies to restrict the activities of the academics, thus reducing competition? Chuck Garber will readily inform you of the meeting that promulagated the rule that says instrumentation purchased with federal dollars cannot be used for commercial purposes. When that rule was made, there was no academic representation (the perspective of scientists at NSF/NIH is different than that of scientists in colleges and universities). Is that fair? What about the politically incorrect white vendor(s) who set up, or use, minority/women fronts in order to sell their products? Is there any noble purpose for vendors who abuse the system for profit and gain? A more important question would ask why is there such a strident militancy against academics among certain individuals at SPI? Let it out, guys, so maybe we can feel your pain......
Chuck
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
In response to the inquiry from anonymous subscriber, ratyrg-at-esvax.dnet.dupont.com, posted on 96-01-24 (regarding availability of cryomicrotomy courses), please be advised that AMC Group offers a series of intensive hands-on workshops on cryomicrotomy on a regular basis. We also offer other advanced workshops including the FIB cross-sectioning and wedge-polishing for site-specific TEM specimen preparation. For the workshops schedule and other informative material on the general subject of specimen preparation, please see our page on WWW at the Microscopy-Online site (http:// www.Microscopy-Online.com/). For further information, please contact me directly. Thanks.
} What about vendors/business people who appeal to the government } and/or granting agencies to restrict the activities of the academics, thus } reducing competition? } Charles J. Butterick (Chuck)
Or, as an actual experience, the SEM vendor who threatened--the correct word--to file a complaint with US Customs, and force a *previous* employer to pay a multi-thousand-dollar penalty if we didn't buy a US-made SEM. (Needless to say, we didn't, and they didn't, since it was a fatuous threat. And a permanent loss of business.) Phil Oshel (Not at U. Ill. at above the time)
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
Group: I am sure that many of you are familiar with the nifty microscopy images produced by David Scharf. If not, allow me to note that they have been featured in Life, Time, National Geographic, Scientific American, Discover, Natural History, Harpers, The Saturday Evening Post, etc. As it happens, I am in the process of producing a series of 16 of his best images - typically as 23 in x 19 in, HIGH quality, lithographic prints. Descriptions, etc. of the series is contained in the next issue of Microscopy Today, which will be mailed in a week's time. Should you, including our international associates, not be receiving the newsletter and have a potiential interest in the prints, kindly advise by eMail and I would be delighted to send you a no cost description of the series. And, should the publication of microscopy prints be an "enjoyable" hobby, I may extend the series. Should you have microscopy images that you consider worthy of reproduction, I would appreciate hear from you. Understanding that there is a bit of "commercialism" in this note, I hope that the uniqueness of the subject causes not many of you to be offended. Best regards to all, Don Grimes, Microscopy Today
Do you know anyone who might be interested in an ISI Super III-A SEM that's in excellent operating condition for only $8,000? We are in the St. Paul, MN area and would be happy to demo the scope in our location on your specimen. This workhorse would be an excellent instrument for a small college/lab/hobbyist or manufacturer. Contact me for more specs or info. Ask me about a $300 travel rebate offer.
} In the past I have used chloroform routinely for expanding thin sections. } Because of the hazardous nature of this chemical I tried a heat pen } (amps - .125), but it was totally ineffective. Would a more powerful } heat pen be effective in reducing compression ? I would appreciate } some feed back on this. } Thanks } Leo
The heat pen I use is fine as long as it is held close to the section and used for around 30 secs (longer for some sections).
Diana van Driel Dept Ophthalmology Sydney University 2006 NSW, AUSTRALIA
X-Sender: mcmouldk-at-uxmail.ust.hk X-Mailer: Windows Eudora Light Version 1.5.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Sixth Asia-Pacific Conference On Electron Microscopy.
Under the Auspices of The International Federation of Societies for Electron Microscopy (IFSEM) and The Committee of Asia-Pacific Societies for Electron Microscopy (CAPSEM).
The Sixth Asia-Pacific Conference on electron Microscope will be held at the Chinese University of Hong Kong, to be held on July 1-5, 1996.
Final call for abstracts.
The deadline for abstracts has been extend to 15th February, 1996. If you are close to the deadline, please use an express mail service to submit your abstracts. Faxed abstracts cannot be accepted.
The conference will provide a forum for the dissemination of new information on the application of scanning and transmission electron microscopy, diffraction and microanalysis in life and materials sciences. The conference will include scientific sessions (plenary lectures, symposia oral presentations and posters). There will also be an exhibition of scientific instruments together with workshops and open laboratories.
******************************************************************* Please contact the above address only. Please do not send requests for information to this e-mail address.
Message-ID: {199602051315.IAA18418-at-IndyNet.indy.net} To: Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
At 02:41 PM 2/4/96 -0500, you wrote:
} Dear Joiner, } If you find a solution to your staining problems, could you please let me } know? I have used the same receipe for UA and LC for over 20 years without } problems. But recently, have been haunted by the symptoms you relayed, with } always using ddwater and trying NaOH washes, different grid suppliers... I am } ready to record the position of the planets when I get good staining. I } wonder if such can be related to low humidity and increased static } electricity???????????? I hope you are able to solve this curse and that you } will share such with me. Needless to say, I will let you know if I have any } luck. } } Best regards, } } Robyn Rufner, Ph.D. } Deborah Research Institute, 20 Pine Mill Rd., Browns Mills, NJ 08015 } 609-893-1016, fax: 609-893-2441, email: rrufner-at-aol.com } ******************** Robyn -
I will indeed share what I've gleened from this thread, but as I told Dwight Beebe, not until you all have solved my problem. No, actually I will pass it on. That's what this listserver is for. The static charge sometimes found on grids has been suggested to be causing precipitation of the stain. If I can find one of those antistatic guns that we used to de-charge our LP's with, I'll try it, being the student of reason and logic that I am and staunch supporter of the scientific meathod. Then at the next full moon I'm going to nail a dead cat to the north side of an oak tree....
Mime-Version: 1.0 Pogany Lajos {pogany-at-power.szfki.kfki.hu} Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
I'm also looking in to CDR's for archiving. I think we will get the Pinnacle 5040, a 2x write 4x read, which runs about $1300 ($1000 internal). HP makes one for about the same cost, Yamaha and Sony have models for a bit more $.
Any comments from people already using any of these?
Is anyone able to assist me in finding a CD ROM writer for image archieving in the price range of about 1000 $? Would be possibile to send me addresses where are those available?
does anyone know if there is a newsgroup or a listserver discussing problems of measuring Ca2+ with fura? Perhaps a group dealing with fluorescence microscopy in general would help.
Thanks for any help
-- Ralf Steinmeyer (Steinmeyer-at-mbox.biophysik.uni-hannover.de) UNI Hannover Herrenhaeuser Str. 2 Inst. f. Biophysik 30419 Hannover
We use Kodak lens cleaner for routine cleaning, plastic dropper bottles are refilled from the more affordable quart containers. Chloroform is reserved for obstinate deposits, like when someone immerses a non-DPX lens into the DPX or Permount.
A couple of microscope technicians that have worked on our equipment use lighter fluid. It dries without leaving a film and yet is claimed not to attack lens adhesives or coatings. Has anyone else used lighter fluid on lenses? Personally, I've only used to take scuff marks of vinyl flooring.
Regards,
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
} Recent messages by Blackwood and Oliver are examples of two opposing points } of view about ethics and philosophy. On one side is the normative view: } that there exist absolute standards ('norms') for conduct. On the other } side is the relative view: standards depend on the situation.
I'm not so sure that it's a question of absolute versus situational ethics as much as *who* defines the "right" and "wrong." My position is not that ethics are situational; my position is that bureaucratic rules and ethics are not the same. Thus, noncompliance is not prima facie evidence that someone has done something "wrong." My primary concern is with the increasing criminalization of noncompliance, leading to allegations of "fraud," "scientific misconduct," and such.
I am not a scofflaw, and I certainly try to live within rules within reason. However, I don't mistake regulatory pronouncements for divine dispensation. Not confusing the two does not equate to the support of anarchy, nor does necessarily make it easy to rationalize any actions, as a different respondent suggests. Indeed, I might suggest the opposite -- that recognizing and accepting individual responsibility for individual ethical decisions independent of an external rulemaking bureaucracy might lead to more careful consideration of action, not less. Relying on legalistic justifications is perhaps not the last, but certainly one of the favorite refuges of a scoundrel.
} Let me remind the reader that we are not talking about human rights } violations, tyranny or murder here. We are only talking about money. It } really isn't worth doing more than 'blowing off steam'. I say follow the } rules that exist and work to change the rules that appear improper.
You bring up another interesting idea, that we are not talking about human rights, but, instead, "We are only talking about money."
In fact, we *are* talking about human rights. It isn't "only money" if a governmental agency, by its regulatory or enforcement rights, deprives you of your livelihood, appropriates your possessions, and criminalizes violation. When investigations result in allegations of fraud there is resultant criminal liability; the only question then is whether or not the regulatory agency wants to take you down bad enough to expend the effort. It may be only money, but when legal fees and seizures destroy life savings it is more than a mere inconvenience. When allegations of fraud or "scientific misconduct" based on technical violations of regulatory pronouncements leads to the destruction of a career, it is a nontrivial life event. For me, at least, it would not be "only money" at risk. As ex-Secretary Donovan said after the trials that destroyed his career in spite of finding him innocent, "Where do I go to get my reputation back?"
When the time and effort it takes to attempt to satisfy the voracious appetitite of a regulatory agency detracts from scientific investigation, patient care, or stifles innovation and experimentation, it is a loss to more than a single investigator. When regulatory demands and limitations force new intellectual exploration and development off the shores of the US, it hurts this nation badly.
In spite of all of this, I would take a more sanguine attitude about today's regulatory atmosphere were it not for the problem that, in my personal opinion, in many areas the regulatory morass is so byzantine and self-contratictory that it is fundamentally impossible to remain in compliance given limited time and resources to devote to doing so. In many a regulatory setting, particularly an investigation, the person being investigated is by no means considered "innocent until proven guilty." Instead, the assumption is that the worst possible interpretation is the correct one, and it is the burden of the person being investigated to prove his or her innocence. And the investigators have all the resources. It is no surprise that even the innocent plea bargain when they face financial ruin in the face of defending themselves against the charges of "fraud," "conspiracy," etc., etc., etc.
In such an atmosphere, simply trying to do one's best is just not good enough. A person who thinks he or she is working within the rules simply doesn't know all the rules. There are no innocent, only the untargeted, and obscurity is a weak defense.
I believe that I am one of the vendors that Chuck Butterick is referring to that offered *free* equipment with the purchase of a certain amount of supplies. It has been a a year or two since I offered the program to Chuck.
He is correct that the idea was to get around budget rules that allow unlimited purchase of supplies but no capital or equipment purchases. In this particular instance, if you bought the equipment and the supplies(film, in this case), paying the lowest available pricing, you would spend the same $ as you would if we gave you the hardware and you paid the premium price for the requisite cases of film.
I will admit that we have a vested interest in placing our equipment with customers so that we sell film. However, I would suggest that if our customers have a need for any of our products and due to various constraints are unable to acquire it through equipment purchase funding, we are working to meet the customers needs-we cant make the customer invest in equipment or participate in a program if they do not want to.
Yes, I am in sales and my livelihood depends on what I sell. I believe that what I sell offers my customers a better way of doing their job;more effectively and more efficiently-if I didnt, I'd quit.
Certainly I would agree that some vendors may have unscrupulous practices and I do not condone that. I am asking you to consider that the intent behind our program and possibly others is to benefit you-so you can have the tools you need to do your job.
John D. Warren Eastern US Sales Manager Helios Scientific Group Polaroid Corporation ______________________________ Reply Separator _________________________________
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. wrote: "......It seems to me that we are all better off if we just force ourselves to live within the rules instead of trying to rationalize our efforts to get around them........".
Andy is correct. The rules and laws must be followed. Those we don't agree with we can work to change. But am I the only one who reads Andy's entire commentary (that only mentions what academics do to get around some regulation in order to purchase an item) and feels resentment? I have been approached many times by vendors who ask me to write lock-out specifications so I will buy from only one source. Other vendors have informed me that if I bought a certain dollar amount of supplies, they would throw in the instrument for free that uses those supplies. More than once I have successfully appealed to the State of Texas on a bid where an unscrupulous vendor misrepresents their instrumentation and/or the price. Some vendors are as eager to earn the academic dollar as the academic is to stretch what little he/she has. What about vendors/business people who appeal to the government and/or granting agencies to restrict the activities of the academics, thus reducing competition? Chuck Garber will readily inform you of the meeting that promulagated the rule that says instrumentation purchased with federal dollars cannot be used for commercial purposes. When that rule was made, there was no academic representation (the perspective of scientists at NSF/NIH is different than that of scientists in colleges and universities). Is that fair? What about the politically incorrect white vendor(s) who set up, or use, minority/women fronts in order to sell their products? Is there any noble purpose for vendors who abuse the system for profit and gain? A more important question would ask why is there such a strident militancy against academics among certain individuals at SPI? Let it out, guys, so maybe we can feel your pain......
Chuck
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
This is a request for information from colleagues experienced with the thinning of TEM specimens held by a Gatan Disc Grinder against South Bay Technology (or 3M Imperial) diamond abrasive films. First some information:
My TEM specimens are taken from joints that have been diffusion bonded by Hot Isostatic Pressing, using the method described in J. Mater. Sci. 29 (1994) 4404-4414. The diffusion couples are as paired semicylinders fitted into a small metal tube (3 mm outer diameter) of stainless steel or nickel. This tube acts as an encapsulation for the isostatic argon pressure (200 MPa) during HIP, and afterwards the capsule is simply sliced with diamond saw before thinning,leaving the capsule as a stabilizing collar around the diffusion couples avoiding cracking for materials having very different coefficients of thermal expansion, such as silicon nitride and Ni-base superalloys. The materials joined so far include Ni-base superalloys (both single crystal and polycrystalline), silicon nitride, titanium nitride, as well as CMC (Si3N4/TiN) and MMC (TiN/Ni) compositionally graded materials (FGM´s).
In June 1995, I participated in a very informative TEM Spec. Prep. Course (lead by Ron Anderson, IBM, East Fishkill Lab, NY, USA) held at BAL-TEC AG in Liechtenstein. After that, I decided to change the type of thinning abrasives from diamond spray on a soft cloth to SBT diamond abrasive films with fixed diamonds (30µm, 15µm, 6µm, 3µm, 1µm and 0.5µm grain sizes), finally followed by colloidal silica on a soft cloth.
However, due to the usefulness of retaining the stabilizing collar of capsule tube, I still want to use the Gatan Disk Grinder (GDG) for parallel thinning, instead of the otherwise impressive SBT Tri-pod polisher demonstrated at the course. Regarding the glue for fixing the specimen on the specimen mount of the GDG, I have changed from wax to Crystalbond 509, in order to avoid remaining films after cleaning in aceton. Finally, I intend to perform low-angle ion beam milling using the BAL-TEC RES 010, possibly preceeded by dimple grinding.
Based on this, I would like your comments on the following questions:
-What are the experiences from the combination Gatan Disc Grinder and South Bay Technology (or 3M Imperial) diamond abrasive films (8´´, plain back), adhering by capillary forces to a slowly (50-100rpm) rotating glass plate?
-Is it always (or exclusively for ductile materials) necessary to use all of the above grain size steps and/or remove a damage layer from the previous step corresponding up to three times the grinding grain size? (If so, given a cutting wheel such as Struers 330CA having 68µm (Grit #220) diamonds, it seems necessary to cut samples thicker than 400µm!)
-Is it necessary/unsuitable to press the GDG against the diamond abrasive film, or is the weight of the GDG itself (660g) sufficient? (If the specimen area would carry the total weight, it would correspond to a pressure of approx. 1MPa (135psi).
-Is it beneficial to rotate the GDG itself during polishing?
-Is it beneficial to grind down the specimen continously or incrementally (50µm steps are recommended by Gatan). If stepwise, should the steps be related to the diamond film grain size?
-Is the fit between the specimen mount and the GDG itself crucial for the integrity of the glued specimen? Which tolerances (after wear) can be tolerated?
Thanks in advance,
/Richard ________________________________________________________________________ Dr. Richard Larker, Ph.D. Phone: +46 920 91107 Research fellow Fax: +46 920 99309 Division of Engineering Materials Lulea University of Technology E-mail: Richard.Larker-at-mb.luth.se S-97187 Lulea, SWEDEN ________________________________________________________________________
I am going to be teaching a mixed TEM/SEM course to undergraduates in Materials Science next quarter. I am sending this note out to ask for comments/suggestions about textbooks, both from people who have taught and those who have been at the recieving end.
NOTE: For obvious reasons, please respond directly to me rather than everyone on the listserver unless you really intend to do the latter.
Message-Id: {9602060920.AA08107-at-smtp-mail.well.ox.ac.uk} X-Sender: levy-at-well.ox.ac.uk X-Mailer: Windows Eudora Version 1.4.4
Please would someone send me the list serv address for subscribing to this list.
Thank you
Elaine Levy -------------------------------------------------------------- Elaine Levy PhD Cytogenetics Laboratory Wellcome Trust Centre For Human Genetics Windmill Rd., Oxford OX3 7BN
tel 01865 740022 fax 01865 742186 email elaine.levy-at-well.ox.ac.uk -----------------------------------------------------------------
our group has received 2 bottles of CIT ALCATEL 220 diffusion pumps oil for free. We have no information at all about how good it would be for our JEOL SEM and we have found no cataloges around. If any of you knows something about vapor presion of this oil, or has experience using it, and has a little time for writing to us, we would appreciate it very much. Thanks in advance,
Silvia Montoro csedax-at-arcride.edu.ar
Regional Center for Research and Development Santa Fe Argentina
} ... recognizing and accepting individual responsibility } for individual ethical decisions ...
To me this seems to be the key; together with a well estabished line of accountability (very important). We will still make mistakes, but hopefully won't be alone in them...
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
I would like to set the record straight about "militancy against academics among certain individuals at SPI" and also, the record with regard to the true complexion of those at NSF who were resonsibile for the drafting and implimentation of NSF Important Notice 91.
First, there is no such "militancy", indeed our firm relies on the graduates of the nation's academic institutions just like any other employer. Our future business prospects in fact are closely intertwined with the future prospects of our academic customers. So any suggestion to the contrary, as they say on the other side of the ocean, is just pure rubbish.
If there is indeed any "militancy" at all, it is on the part of a small handfull of individuals and departments who are literally running businesses out of their university laboratories and who feel threatened should anyone suggest that what they are doing is not right.
So surely there is concern about anyone using university equipment in competition against private sector laboratories such as our own. For the most part, people I know in academia are on "our" side of this issue and hold the view that the university's instrumentation in fact should not be used this way. They seem to feel that those projects that are basic and fundamental in nature and suitable for inclusion in a student's thesis (e.g. those projects that do contribute to educational objectives) take up the available time for their instrumentation anyhow and that outside commercial work just gets in the way and interferes with the progress of the students and the quality of the work being done. So those who advocate some contrary view are clearly on the wrong side of the tracks on this issue, and are also on the wrong side with respect to their own academic peers.
Second, at the time of the drafting of NSF Important Notice 91, the Director of NSF was Prof. George Pimentel, perhaps one of the most respected and published academicians ever to head NSF. He came to NSF from the Chemistry Department at the University of California-Berkeley. He was an "academic" and represented "academic interests" in every sense of the word. To suggest otherwise is to misrepresent Dr. Pimentel's attitudes, perspectives, and outlook.
The Associate Direrctor at the time was Dr. Donald Langenberg. Prior to his position at NSF he was a full professor of physics (at the U. of Pennsylvania, if I remember correctly), and after his tenure at NSF moved on to serve as President of the American Association for the Advancement of Science, the Chancellor (I think that was the postion) of the University of Illinois - Chicago and from there to his present position of Chancellor of the University of Maryland system, with over sight over all state universities in the State of Maryland.
Both Drs. Pimentel and Langenberg were in attendence at most of the meetings that led up to the drafting of NSF Important Notice 91. And their participation in the drafting of the document was certainly in no way to be thought of as being passive. So for anyone to suggest that in some way the academic community was not properly represented is again "pure rubbish".
But intentional or not, it is obvious that there are still some who would try to discredit the NSF Important Notice 91 document in every possible way, in this case, by suggesting there was a lack of academic representation among those involved with its drafting and implimentation.
With regard to some of Chuck Butterick's other comments, unethical behavior of any type should not be tolerated in any environment where high quality research is being conducted. Vendors guilty of the kinds of infractions described in Chuck's posting should be reported to the appropriate procurement officials who have in place mechanisms by which such abuses of process can be rooted-out and stopped. Of course it does take two to tango and therefore the other side of the equation, that is, the customer who is encouraging such inappropriate practices should also be reported. While the wheel of "justice" might move slowly, the wheel does move, but please don't paint the behavior of the entire vendor community with the same brush. The electron optics industry of manufacturers and distributors by and large is highly professional and honest and when instances do occur that do not fit this pattern, there is a process by which one can keep that kind of circumstance from recurring in the future.
As a final comment, for anyone not personally familiar with NSF Important Notice 91, I would be happy to FAX you a copy, just send me your FAX number.
Chuck (Garber) ====================================================== 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: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
At 08:21 AM 2/6/96 EST, you wrote: } Joiner- } If you *do* find an old Zerostat gun (which is a little like searching for } brontosaurus chow), it's probably not going to be any good any more. The same } effect can be acheived by using glow discharge. Have you got that on an old } evaporator or coater? Good luck! } Steven } } ************** Yes, I do; although the evaporator ain't no spring chicken.
Message-Id: {199602061557.JAA23186-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 09:06 AM 2/6/96 -0500, you wrote: } Good Morning. } } Further to the note I sent you last week where I fingered the grids as my } most likely suspect for the introduction of Pb hexagons: } } My understanding is that these things are made by a plating method. We had a } program here for a while looking at the microstructure of thin film magnetic } media for one of our then subsidiaries. The magnetic media is deposited on } plated substrates. } } One of the things we struggled with, is that the plating procedure was done } in baths contained by tanks soldered together with (you guessed it) Pb. The } Pb impurities found their way into the plated metal (concentrating at the } surface), and when I would microtome these things, I'd get hexagons aplenty! } } Perhaps the source of contamination in the grids? }
********************* That has been proposed (the grids being the source) and we are gearing up to improve our grid washing.
On June 22, 1995, Chuck Garber wrote, "A special committee, made up of top NSF officials (the late Dr. George Pimentel played a key role in the formulation of the final document as did also Dr. Donald Langenberg, now Chancellor of the University of Maryland system), lawyers representing the NSF (Mr. Charles Herz who is still at NSF in that same capacity), members of the independent laboratory community (including myself) plus some other knowledgeable people came up with the first draft of NSF Important Notice 91."
I still stand by my statement, no academic representation existed. Chuck Garber was once a part of academics, but he doesn't represent academic interests. NSF scientists, though academics at one time, are representing NSF and no one else. That committee appears to have been a stacked deck of cards. This is a democracy. Bad law can be overturned.
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
Low temrerature embedding/infiltration is not an easy procedure. The concetration and the distribution of the soluble proteins play an important role of successful LT embedding. I have had some success to retain soluble proteins in different type of plant tissues. Enzyme activity in vacuoles and in cell wall,using post-embedding techniques is our interest. Lowicryl resins: HM23 and K4M, as well as LR White were used at different temperatures and times (-80 to -20C; 2days to 2 weeks) after "mild" GA fixation OR cryo- fixed plant tissues, using PLT techniques (dehydration at progressively lower temperatures). LR White, after UV polymerization developed very small air (?) "packets". It doesn't section happily, yet not a major problem. "An Osmium-free Method of Epon Embedment That Preserves both Ultrastructure and Antigenicity for Post-embedding Immunocytochemistry", Phen, K.D., Rustioni.A., and Weinberg, R.J., J. Histochem.Cytochem. 42, 1995. You may adopt this procedure to LR White. It should work well. Good luck! Any other ideas, please let me know. Regards,
Laszlo J. Veto Electron Microscopist Agriculture and Agri-Food Canada Summerland, BC Ph: 604-494-7711 Fax: 604-494-0755 e-Mail: veto-at-bcrssu.agr.ca
} Robyn - } } I will indeed share what I've gleened from this thread, but as I told Dwight } Beebe, not until you all have solved my problem. No, actually I will pass it } on. That's what this listserver is for. The static charge sometimes found on } grids has been suggested to be causing precipitation of the stain. If I can } find one of those antistatic guns that we used to de-charge our LP's with, } I'll try it, being the student of reason and logic that I am and staunch } supporter of the scientific meathod. Then at the next full moon I'm going to } nail a dead cat to the north side of an oak tree.... } } * * Joiner Cartwright, Jr. * *
Barring a Zero-Stat gun, which I've still seen advertised somewhere, try touching your charged grid to a bit a metal on a grounded piece of electrical equipment. It's safe (this is how some powerbars drain your personal static charge so you don't zap your computer). Phil
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
I received a used ISI DS-130 SEM equipped with a cathodoluminescence (CL) detector. This detector has no ellipsoidal mirror coupling to it. I guess this must be a "diode" detector which can detect CL directly from the sample.
(1) Does anyone have information about this type of detector ? How does it compare to the detector with an ellipsoidal mirror ?
(2) Does anyone have experience taking CL images of quartz? I know we cannot focus the CL, but at least we can use proper settings (by adjusting accelerating voltage, beam current, etc) to optimize the CL resolution. Thanks.
Long Liang ARCO EPMA/SEM Lab lliang-at-is.arco.com
} On June 22, 1995, Chuck Garber wrote, "A special committee, made up } of top NSF officials (the late Dr. George Pimentel played a key role in the } formulation of the final document as did also Dr. Donald Langenberg, now } Chancellor of the University of Maryland system), lawyers representing the } NSF (Mr. Charles Herz who is still at NSF in that same capacity), members } of the independent laboratory community (including myself) plus some other } knowledgeable people came up with the first draft of NSF Important Notice } 91." } } I still stand by my statement, no academic representation existed. } Chuck Garber was once a part of academics, but he doesn't represent } academic interests. NSF scientists, though academics at one time, are } representing NSF and no one else. That committee appears to have been a } stacked deck of cards. This is a democracy. Bad law can be overturned. } } } } } Charles J. Butterick (Chuck) } Electron Microscopy Center } Department of Cell Biology } and Biochemistry } Texas Tech University Health } Sciences Center } 3601 4th Street } Lubbock, Texas 79430 } } vox (806) 743-1633 } fax (806) 743-1219 } email emccjb-at-ttuhsc.edu or } chuck-at-micron1.lubb.ttuhsc.edu
Dear Chuck's (Butterick and Garber),
I really believe that this sort of dialog (?!) does not belong here.
Thank you for the excitement, though!!!
Best regards
Massimo Sassaroli, D.Sc. Dept. of Physiology & Biophysics Box 1218 Mount Sinai School of Medicine 1 Gustave L. Levy Pl. New York, NY 10029-6574
We would like to know if anyone has an opinion on the effectiveness of using a silver screen and double projectors for 3D imaging. We do not want to commit ourselves to buying a screen (which is quite expensive) and then find it has been a waste of money.
We intend to use the system for illustrating talks to members of the the public visiting the Unit and also for student classes. We would be projecting mainly SEM images.
We would like to hear from anyone who has tried this or any other alternative method of displaying 3D images to groups of people.
Thanks in advance,
Mark Gould
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
I am seeking facilities for SEM-EDS and/or SEM-WDS in proximity to Berkshire county, Massachusetts (to include western MA, southern VT and eastern NY). Anyone knowing of a commercial, industrial or academic facility in this area, please contact me directly.
Dear Dr. Hendriks, (and other colleagues interested in the topic related to the first question (Q1) below),
I have the following comments/questions (C1-6) caused by your answers (A1-6) to my questions (Q1-6):
Q1: What are the experiences from the combination Gatan Disc Grinder and SouthBay Technology (or 3M Imperial) diamond abrasive films (8´´, plain back), adhering by capillary forces to a slowly (50-100rpm) rotating glass plate?
A1: While I have not used the Gatan tool myself, I can answer you in terms of using our SBT Polishing Fixtures. Other than the Tripod Polisher (R) we also have other fixtures which are similar in size and weight to the Gatan. We have had very good experience in having the films remain firmly attached - which I presume is your question. We typically recommend that people use the plain back films as they adhere well, are easy to remove without damaging them and then can be re-used. This is important as the films are not inexpensive! You may also want to note that the bottom of the Gatan fixture is stainless steel (I believe) and it will wear when polishing with diamond. The SBT fixtures described above have tungsten carbide feet at the bottom to help resist wear. However, even the tungsten carbide will wear when polishing diamond. For final polishing on samples less than a few microns thick, it is best to use a much slower wheel - ie less than 5 rpm.
Comment 1: In my tests so far, the SBT diamond films were firmly attached to the wet glass plate by squeezing the water out simply by pressing a PMMA ruler in several directions on the films. In spite of this, the specimen (glued to the mount by Crystalbond 509 at 130 C) sticked several times to the 30 micron diamond film and loosened from the mount, causing expensive scratches in the film! I tried to reduce both the incremental steps (from initially 25 micron down to 5 micron steps) and the rotational speed from 100 rpm to 50 rpm, but with limited success. I experienced the same problem with the 6 micron diamond film (but not with the intermediate 15 micron!) when using 2.5 micron steps. The feeling during this "seizure" was that the whole GDG was sucked (by capillary forces?) firmly to the film, twisting it loose, causing scratches and loosening of the specimen from the mount. This kind of experiences did not occur earlier when using diamond spray on soft cloths! Perhaps I have used a little too high pressure on the GDG, but the unpleasant tendency for it to "follow" the film was noticeable even without any extra load. It was even noticeable when trying without any specimen mount present in the middle of the GDG! This was peculiar, since the instructions inside the cover of the Gatan Disc Grinder 623 specify the following: "Polish specimen preferably using 3M Imperial Lapping films". Furthermore, the instructions recommend as large increments as 50 micron! Concerning your comment that the WC-Co feet of the SBT fixtures should wear less than the large SS plate of the GDG does; supposing that is true, what does it imply for the wear of the soft Delrin feet of the Tripod? How is it then possible to know that the micrometer is not "underestimating" the thinning of the specimen? What is the influence of the specimen/feet area ratio?
Q2: Is it always (or exclusively for ductile materials) necessary to use all of the above grain size steps and/or remove a damage layer from the previous step corresponding up to three times the grinding grain size? (If so, given a cutting wheel such as Struers 330CA having 68 micron (Grit #220) diamonds, it seems necessary to cut samples thicker than 400 micron!)
A2: The short answer is NO. You do not need to use every grain size. You do however, need to be mindful of the 3x rule when polishing. I assume that you are interested in the microstructure of the sample being polished. You can polish a sample from 500 microns thick down to 5 microns thick by using only 0.5 micron film. The problem is that it will take you many, many, many hours and consume many pieces of film. Reason the many steps are used is to minimize the polishing time. You can obviously remove material much faster with a 30 micron film than with a 1 micron film, but you will also cause more damage. The polishing sequence is determined by 1) amount of material to be removed 2) desired final thickness 3) budget for film 4) level of patience. I do not fully understand your example of the Struers 330CA so I won't address that here.
C2: The example of the Struers 330CA diamond cutting wheel, which I have used for slicing my samples up to now, was after some investigation found to have a diamonds of 68 micron size (Grit #220), and if this causes 3x68 micron of damage on each side, and you follow the 3x rule with 30-15-6-3-1-0.5 micron diamond films, you simply need to cut thicker (} 740 micron!) than I have been doing so far (400 micron) to end up with any specimen thickness at all! However, may it be the case that the cutting wheel causes less than the anticipated 3x diamond grain size damage, due to the low level of normal force against the surface during cutting, when compared to polishing conditions? Regarding thinning of ceramics like silicon nitride, which diamond grain size steps might be omitted ? Can any step be omitted for Ni-base superalloys?
Q3: Is it necessary/unsuitable to press the GDG against the diamond abrasive film, or is the weight of the GDG itself (660g) sufficient? (If the specimen area would carry the total weight, it would correspond to a pressure of approx. 1MPa (135psi)).
A3: In general, it may be advisable to add some additional pressure during the rough grinding stages, however, the during final polishing stages it is best to minimize the load on the sample. Unfortunately, the Gatan Disc Grinder has no facility which will allow you to reduce the specimen load below the weight of the fixture. If you have had problems with samples cracking when thinning at the final stages, excessive load is probably the problem.
C3: Due to the experiences given in C1 above, I am not confident with adding any extra pressure at all! Regarding the thickness of the water film between the GDG and the diamond film, are there any estimations of the "regimes of lubrication" (hydrodynamic, partial or boundary lubrication, as related to the diamond grain size) below the GDG or SBT polishing fixtures, or even below the specimen itself?
Q4: Is it beneficial to rotate the disc grinder itself during polishing?
A4: Yes, for several reasons. It is best to continually introduce new cutting faces will increase removal efficiency and also reduce preferential scratching in the surface. However, in some instances, people will continue polishing in 1 orientation per grit size. This will provide a scratch pattern in one direction. When changing to the next size film, you can rotate the sample 90 degrees and see a new scratch pattern develop. Once the new scratch pattern covers the surface, you know that you have removed the scratches from the previous grit size - but not the damage! Remember the 3x rule.
C4: For the GDG, the specimen mount may not be unambigously prevented from rotating relative to the GDG orientation; is this also the case for the SBT polishing fixtures that you have described?
Q5: Is it beneficial to grind down the specimen continuously or incrementally?
A5: This is a question which is peculiar to the Gatan Disc Grinder. With the Gatan Disc Grinder your specimen is mounted to a post which DOES NOT freely float inside the larger outside ring. Polishing is accomplished by extending the sample below the surface of the larger outside ring. This means that the entire weight of you fixture is resting on your sample. There is a great likelihood that you will damage your sample and/or round the edges of your sample. Because of this inadequacy, Gatan recommends an incremental procedure which minimizes the effect, but maximizes the effort! In more thoughtful designs (such as the South Bay Technology Fixtures :-)) the sample is mounted to a free floating rod which is gravity fed into the abrasive surface. The load on the specimen is thus spread across the entire base of the fixture. Edge rounding is eliminated because the edges of the sample are protected by the base of the fixture. Using this type of fixture, the incremental polishing is not necessary.
C5: How are the SBT polishing fixtures designed to stop further thinning of the specimen after reaching the intended thickness in each step?
Q6: Is the fit between the specimen mount and the disc grinder crucial for the integrity of the glued specimen? What tolerances can be tolerated?
A6: Think of the disc grinder as a 2 part device 1) the piston with a sample mounted at the bottom 2) an outside ring with a center hole. The fit between the piston and the center hole should be very precise. This precision is increased by making a longer contact area between the 2. Unfortunately, the Gatan Disc Grinder has a very short contact area which will magnify any problems. As the Gatan piston/hole are nearly in contact with the abrasive surface, I would think the chance for abrasive wearing away the center hole is rather likely. The sample mount is replaceable so that part is not a big problem. I would think that if you do wear the center hole uniformly, you could make special sample mounts to correspond to the new center hole size. Certainly the accuracy in these 2 areas is crucial - especially when polishing very thin. I did notice that you are familiar with Tripod Polishing and that you said you did not want to do that because you wanted to retain the stabilizing collar. Since you attended Ron Anderson's course, he probably spoke to you primarily about "wedge polishing". You may like to know that it is also possible to use the Tripod Polisher (R) for parallel polishing by mounting the sample in the center of the 3 micrometers. In fact, this was the way Tripod Polishing originated. The "L" brackets and wedge polishing techniques were later developments. You also mentioned that you now use Crystalbond 509 for mounting your sample to the holder. As a matter of interest, we supply an identical product called QuickStick 135 which can be purchased in a package of 20 small (3" x .25" x .25") unwrapped sticks under part no. MWH135. An alternative may be to use "super glue" or any cyanoacrylate. It bonds quickly and removes completely in acetone. The only problem is that removing it is somewhat unpredictable. Sometimes it can come off quickly, other times it may take quite some time. It is also important to be sure to use fresh super glue. We use super glue for wedge polishing of our TEM samples. You can pick it up at a local hardware store. If you would like information on any of our products, you can contact me directly or you may contact our office in Sweden, Tudor Barnard (snip)
C6"+": How useful is the Tripod for parallel (not wedge) polishing, compared to the other SBT polishing fixtures that you have described? Regarding your representative in Sweden, Tudor Barnard, I bought my SBT diamond films through him, but he was at that time (august ´95) not able to find a cyanoacrylic glue that was soluble in acetone (but only in di-methylformamide or tetra-chlormethane, which I consider to be far more harmful substances!)
Thanks in advance,
/Richard ________________________________________________________________________ Dr. Richard Larker, Ph.D. Phone: +46 920 91107 Research fellow Fax: +46 920 99309 Division of Engineering Materials Lulea University of Technology E-mail: Richard.Larker-at-mb.luth.se S-97187 Lulea, SWEDEN ________________________________________________________________________
We have used the silver screen method of projecting 3-D slides of all varieties for over 10 years. We find it by far the best method for displaying 3-D material to a large group of people. We use separate projectors with cross polarizers (I have heard of but not seen large screen projector wich takes a single slide containing two images).The down side of the procedure is the time it takes to align the two projectors and the necessity that all slides be exactly aligned in the same way. Nothing will make an audience sea sick faster than realigning projectors between stereo-pairs to accomodate different alignments. We sometime spend an entire day making sure a group of slides is all set up similarly. For this reason, for showing one or two people our stereo work we use premounted pairs in small, hand held viewers. However, this is not nearly as dramatic as a huge image on a screen. Any one who has ever stood in front of an audience and presented a stereo presentation will also tell you that the second down side of the method is trying to keep your composure while looking out at an audience full of people wearing silly dark glasses in darkened room.
I hope this helps-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Wed, 7 Feb 1996, Richard Easingwood wrote:
} } We would like to know if anyone has an opinion on the effectiveness of } using a silver screen and double projectors for 3D imaging. We do not want } to commit ourselves to buying a screen (which is quite expensive) and then } find it has been a waste of money. } } We intend to use the system for illustrating talks to members of the the } public visiting the Unit and also for student classes. We would be } projecting mainly SEM images. } } We would like to hear from anyone who has tried this or any other } alternative method of displaying 3D images to groups of people. } } Thanks in advance, } } Mark Gould } } } Richard Easingwood } South Campus Electron Microscope Unit } Otago Medical School } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } SOUTHERNMOST E.M UNIT IN THE WORLD } } } } } } } } } } } }
+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+ Simone T. Boesch {simone.t.boesch-at-uibk.ac.at} Dept. of Zoology, University of Innsbruck Technikerstrasse 25, A - 6020 Innsbruck, Austria phone: (43)-512-507-6171, fax: (43)-512-507-2930 +-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+
A simple way of projecting stereo images is to get a friendly photographer to double expose your stereo pair through red and green filters onto slide film. All you need then is to hand out the old red green glasses and appeal for them to be returned! Its cheaper that way!
The other main item that you need to know is that there is a convention as to which way around the exposures are made so as to register re. the red and green and whch way around the glasses are held.
Keith Ryan Plymouth Marine Lab. Citadel Hill Plymouth PL1 2PB, England
Does anybody know if Gatan has a WWW-site with FAQ´s on the net, or even just an E-mail address?
Thanks in advance,
/Richard ________________________________________________________________________ Dr. Richard Larker, Ph.D. Phone: +46 920 91107 Research fellow Fax: +46 920 99309 Division of Engineering Materials Lulea University of Technology E-mail: Richard.Larker-at-mb.luth.se S-97187 Lulea, SWEDEN ________________________________________________________________________
We are encountering problems in disposal of uranium salts as our site safety officer considers it as a mixed/radioactive waste. I wonder what is the experience of other microscopists regarding radioactivity/disposal of uranium compounds.
A student in Chemical Engineering wants to examine pore membranes in the SEM. Currently we are looking at the membrane surfaces but he was wondering about viewing the membranes in cross-section. Does anyone know a method to prepare such a section? Please respond to his email address at the University of Missouri at c517837-at-showme.missouri.edu or you can contact me directly.
Thanks in advance.
Lou Ross
101 Geological Sciences Bldg. University of Missouri Columbia, MO 65211 (314) 882-4777, 882=5458 fax
We have recived a JEOL 100B electron microscope (Series 155037-30) as a gift from University of Pennsylvania. Although this microscope is in good conditions, it needs complete rehabilitacion. We have highly qualified personnel to do this job but we found that the instruction manual and circuits diagrams we have recived, do not much with the specifications of the microscope. If somebody has these manuals from JEOL 100B electron microscope (with the series number higher than ours) which is not longer useful, we can do good use of them in this part of the third world.
Dra Patricia Pons Centro de Microscopia Electronica Universidad Nacional de Cordoba Cordoba - Argentina
Does anyone have information on TMAH as a back-etch for silicon devices? I have one paper "A New Back-Etch for Silicon Devices" P. Malberti, M. Ciappa , P. Scacco. Also, in this paper it refers to a "bain marie". Can anyone tell me exactly what it is? I am assuming it is like a double boiler.
Terri Mengelt Motorola COM 1 Phoenix, Arizona 85201
I would like to thank everyone who responded to my earlier email (see the end). A number of you were interested in the responses, which I am including here (with editorial comments omitted). As a follow up, I heard almost exclusively from faculty and only one student - how about a few more comments from the students (I will preseverse anonimity).
This is a list of the responses:
4 Responses suggesting (but SEM only) "Scanning Electron Microscopy and X-Ray Microanalysis", by Goldstein, et.al., Plenum Press
3 Responses suggesting "Electron microscopy and analysis" P.J.Goodhew and F.J.Humphries, Taylor and Francis 1988.
"Light and Electron Microscopy" by Slaytor and Slaytor.
"Scanning Electron Microscopy and X-Ray Microanalysis" by Robert E. Lee
"Scanning and Transmission Electron Microscopy," by Flegler, Heckman and Klomparens (W.H. Freeman and Co., 1993)
"Electron Beam Analysis of Materials" by Mike Loretto
---- Copy of Original Message -----
I am going to be teaching a mixed TEM/SEM course to undergraduates in Materials Science next quarter. I am sending this note out to ask for comments/suggestions about textbooks, both from people who have taught and those who have been at the recieving end.
NOTE: For obvious reasons, please respond directly to me rather than everyone on the listserver unless you really intend to do the latter.
We've got a minor glitch in the subscribe/unsubscribe system. Unfortunately, I'm on vacation and too far away to fix the link. This just means there will be a delay if your trying to unsubscribe. I'll be logging in remotely and doing manual updates, but the connections are sometimes slow from here.
I'll be back in town the middle of next week and we should be back to normal.
Sorry for the inconvenience
Nestor Your Friendly Neighborhood SysOp
Weather report: 34 C, partly cloudy, and the view on the beach is great. ;-)
Message-Id: {199602071600.KAA28036-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 05:46 PM 2/6/96 -0500, you wrote:
} Dear Chuck's (Butterick and Garber), } } I really believe that this sort of dialog (?!) does not belong here. } } Thank you for the excitement, though!!! } } Best regards } } Massimo Sassaroli, D.Sc. } ********************************
Massimo -
At the risk of adding UNNECESSARY fuel to the fire, I'm going to disagree with you. These are issues that are indeed important, and if these two gentlemen (and they ARE gentlemen) can hash them out without resorting to nastiness, we will all benefit. It is easy for us academics to overlook the position of the commercial/for profit operator, and I assume that the converse is also true.
I would point out that there may be more than two sides to the question, or at least an intermediate position. My lab, for example, is in a medical school and I work WITH investigators who pay me for my services with their grant money. My lab receives no direct grant funding, either in its capital set-up and improvements or its operation. The only financial support we get is what we earn in fee-for-service.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
I am a microscopist with 21 years experience in Scanning Electron Microscopy in materials science. I am seeking a new position as a lab supervisor or a lab support technician. If you have or know of a position available that would utilize these basic qualifications, and to receive a copy of my resume, please contact me:
We've got a minor glitch in the subscribe/unsubscribe system. Unfortunately, I'm on vacation and too far away to fix the link. This just means there will be a delay if your trying to unsubscribe. I'll be logging in remotely and doing manual updates, but the connections are sometimes slow from here.
I'll be back in town the middle of next week and we should be back to normal.
Sorry for the inconvenience
Nestor Your Friendly Neighborhood SysOp
Weather report: 34 C, partly cloudy, and the view on the beach is great. ;-)
Please publish the following information in the MicroWorld e-mail newsletter:
1996 Lehigh Microscopy Short Courses June 10-21, 1996 SEM, X-ray Microanalysis, AFM and SPM, AEM Contact: Sharon L. Coe Department of Materials Science & Engineering Whitaker Labaoratory 5 East Packer Avenue Bethlehem, PA 18015 610/758-5133 (phone) 610/758-4244 (fax) slc6-at-lehigh.edu http://www.lehigh.edu/~inmatsci/Microscourses.html
Thanks you for your help.
Sharon L. Coe Department of Materials Science & Engineering 5 East Packer Avenue Lehigh University Bethlehem, PA 18015 610/758-5133 e-mail: slc6-at-lehigh.edu
I have projected stereo images successfully by creating red/green anaglyphs on transparency material. We took our digital images, combined them using PhotoShop or similar program, and printed them on a dye-sublimation printer. Using a standard overhead projector and the red/green glasses, the images greatly impressed my audience. The dual projector system seems a bit specialized.
Henk
} We would like to know if anyone has an opinion on the effectiveness of } using a silver screen and double projectors for 3D imaging. We do not want } to commit ourselves to buying a screen (which is quite expensive) and then } find it has been a waste of money. } } We intend to use the system for illustrating talks to members of the the } public visiting the Unit and also for student classes. We would be } projecting mainly SEM images. } } We would like to hear from anyone who has tried this or any other } alternative method of displaying 3D images to groups of people. } } Thanks in advance, } } Mark Gould } } } Richard Easingwood } South Campus Electron Microscope Unit } Otago Medical School } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } SOUTHERNMOST E.M UNIT IN THE WORLD } } } } } } } } }
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- An optimist believes that we live in the best of all possible worlds. A pessimist fears this is true.
} I have projected stereo images successfully by creating red/green anaglyphs } on transparency material. We took our digital images, combined them using } PhotoShop or similar program, and printed them on a dye-sublimation } printer. Using a standard overhead projector and the red/green glasses, } the images greatly impressed my audience. The dual projector system seems } a bit specialized. } } Henk
The Anaglyph system is indeed simple and easy to use and requires only an ordinary projector and screen but it does have at least two snags. Apart from the opportunities that it gives to "Murphy" to attack when two exposures must be recorded on the same piece of film in register and through different colored filters, many people get very uncomfortable when the color of the image presented to one eye is very different from that presented to the other. The phenomenon is referred to as "color bombardment" and was much discussed by Vernon Barber in the late seventies/early eighties with reference to Scanning Electron Microscopy.
The second snag is that you cannot show colored images using this system. For instance you cannot use it to show stereo views of confocal data in which you have superimposed separate 3D images obtained from 2 or 3 different fluorescent dyes in the same specimen. Likewise, you cannot show stereo SEM images in which the SE signal is recorded in tones of grey while the BSE image is superimposed in shades of red.
In addition, although there is usually less overlap between the red and blue filters than between the red and the green, the light output in the blue from most projectors is insufficient to convey many grey levels to the screen and back.
Once you accept the difficulty of finding an "aluminized" screen, the Pol system has much to recommend it.
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
I have done just as you suggest. Using a portable silver lenticular (sp?) screen (~$200.00) which does not depolarize the images, a beautiful 3D image slide show can be projected.
In our department, we have had the capability of mixing 2D and 3D images in a single slide presentation for about 5 years now and I have even done a few 3D demonstrations at a local high school -- with great success (held their attention from start to finish)!
As you have outlined, two matched projectors are required as well as polaroid transparancy sheets and glasses. Using a single piece of Scotch Tape to attach (hang) a piece of the polaroid transparancy over the front of the objective lens of each slide projector (each piece oriented 90 degrees to the other) is sufficient to project a polarized image on the screen.
The glasses, which each person viewing the projected images must have, represents a bit of work. I made a template for the eyes (no ear supports/hangers needed - see below dowell sticks) and traced the template image for as many glasses as I needed on firm, but flexible, cardboard that could be cut with a good pair of sharp tipped sissors.
After tracing and cutting the cardboard, using carefully applied small pieces of Scotch Tape, I taped the polaroid 'lens pieces' from the same sheet of polaroid material used in front of the projector lenses [CORRECTLY ORIENTED] for left and right eye. By correctly oriented, I referring to the necessity of having all glasses with the angular orientation of the polaroid pieces identical -- all left and right eye orientation must be the same in the glasses and matched to the orientation of the pieces in front of the projector lenses. This way, all observers receive the same (correct) image in the left and right eye.
When the 'lenses' were all taped in place, I then taped these glasses by their right or left edge (makes no difference) to a 12 inch length of 1/4 inch wooden dowell stick so that viewers could hold the glasses in front of their eyes (and personal prescription glasses if that's the case) much like holding those small masks at a masquerade party. This may seem a little amusing, and sometimes gets a laugh at the beginning of a 3D presentation, but it was the method I developed to circumvent any health hazard concerns from the repeated public use these glasses -- this is why there are no ear supports as on regular glasses.
From the SEM image standpoint, I take two photos of the same subject (carefully aligned to be nearly the same field) at angles of +/- 5 degrees off verticle by stage tilts. I can then make direct positive slide transparancies with B/W 35 mm microfilm for projection. Both images are matched for size when projected on the screen and simply superimposed -- the glasses and polaroid pieces do all the separation of images to create a very clear and satisfactory 3D image for the viewers.
Hope this helps. This note has been a sort of 'stream of thought' description so please don't hesitate to ask any questions.
} We are encountering problems in disposal of uranium salts as our site } safety officer considers it as a mixed/radioactive waste. I wonder } what is the experience of other microscopists regarding } radioactivity/disposal of uranium compounds. } Dear Manoj, The law in New York allows us to pour uranyl acetate down the regular sink for the amounts used for staining. For larger amounts, they have speci- fied procedures, and our responsibility is to give the material to them with the activity listed on a data sheet--they do the rest. Yours, Bill Tivol
In response to you questions regarding the use of a GATAN disc grinder on diamond lapping films:
While I don't mean for this response to be an advertisement for product, I must preface my remarks by mentioning that the experiences and methods described here occurred using the products of my employer: BUEHLER, LTD in our development laboratory. Therefore, I, like Mr. Henriks, do have a commercial interest in your successful specimen preparation.
A1: In response to your query on experiences with GATAN's disc grinder on Diamond Lapping Films, I also have not used the GATAN fixture; however, I have tried (successfully, I might add) to grind metallurgically mounted samples (1-1/4" in diameter) on BUEHLER's ULTRA-PREP diamond films. The problem of adhesion between the sample and the film, when water is used as a lubricant, does cause movement of the diamond film on the glass wheel. Our experiences indicate that wheel speed makes no difference. However, success was achieved by using a PSA (Pressure Sensitive Adhesive) backed film, attached to the surface of a POLIMET Pad. It was determined that the diamond films are extremely flat compared to comparably sized silicon carbide papers, and therefore a thin film of water between sample and abrasive film causes the sticking problem. Breaking up this film of water is the key to stopping the sticking. The POLIMET pad is perforated, leaving small depressions in spots under the film, while maintaining the rigidity needed to keep the sample flat. Another option for you is to break up the smooth, flat surface in contact with the film; i.e. the bottom of your GATAN fixture (this may not be the more convenient option, however).
With regard to your question about the DELRIN feet of tripodding type polishers, I have had experience here as well. BUEHLER's TRIPOINT POLISHER(TM) also uses DELRIN feet. These DO wear when held against the diamond films. However, the purpose of the TRIPOINT POLISHER(TM) type fixtures was originally to cross-section microelectronic materials. While the DELRIN does abrade slowly, it does not abrade anywhere near the rate of silicon, GaAs, etc. The micrometers on which the feet are mounted are there for the purpose of adjusting tilt and pitch of the sample, as well as adjusting for the insignificant (when polishing most materials) wear encountered on the feet.
The influence of the DELRIN foot area is important with regard to the sticking problem. Obviously, the smaller the feet, the less sticking that will occur. However, another variable should be considered: The abrasive size. On a 30 micron diamond film, the TRIPOINT POLISHER(TM) does not stick because the diamond is large enough to allow air to break up the water film between foot and diamond film. However, when you get in the smaller sizes, e.g. 1 to 0.5 microns, the DELRIN feet become 'polished', and the sticking problem begins to increase. This is sometimes felt as a steadily growing chattering of sample on abrasive surface. My suggestion is to cut grooves in the DELRIN feet in order to break up the large surface area found there.
A2: The 3X rule of abrasive damage removal was proposed by a competing metallurgical supply company a number of years ago. It is a very good guideline to work with, but it is not a scientifically proven rule, as far as I am aware. Of course ductile materials will tend to follow the rule more than hard, tough materials. Therefore, I don't see any reason why you must use every abrasive size while grinding your materials. However, for samples where edge retention is important, or where a cross-section of extremely thin layers is being attempted, I have found that the prescribed steps of 30, 15, 6, 3, 1 and 0.5 micron diamond films are necessary. Especially the latter case.
If you would like to eliminate a significant portion of the grinding that is necessary, you may want to try a diamond wafering blade which contains a finer abrasive than the 68 micron diamond you are currently using. We offer a blade which contains diamond about 1/13th this size, and which is designed for exactly this purpose. We call this the L5 series blade. You can contact me directly by e-mail, phone or fax for more info. The surface finish produced is close to that produced by a 3 micron abrasive film. I do have a BUEHLER produced, technical brochure (Note: contains info on only our products) which describes the effects of cutting force, speed, abrasive size, etc., if you would be interested.
A3: Our experience has been that low loads give better results on diamond lapping films than high loads. This is especially true with ductile materials. Cleaning the films with a fresh, folded paper towel during the grinding step also improves the result obtained on most materials. The towel captures loose diamond and swarf before it can be reintroduced to the sample; causing scratches or embedding. Make sure to use a fresh towel with every abrasive size change.
A4: I differ with Mr. Henriks on this question. I don't suggest rotating the specimen during each grinding step. Rotating the specimen does not introduce fresh abrasive to the sample. Only the wheel rotation does this.
When the scratch pattern from the previous abrasive is removed, the damage may or may not be removed (again, depending on the material). However, this may not be relevant in the case of ceramics such as the silicon nitride you mentioned. This is because the damage mechanisms are different between ductile and hard/tough materials.
A5: I do agree with Mr. Henriks on this question. I was, at one time, Application Engineer at SBT, and have had the opportunity to work with all of their carbide footed grinding fixtures. The free-floating nature of their fixtures does reduce the load on the specimen. I believe they still offer a fixture with spring counterbalancing, which further reduces the specimen load. These fixtures have an adjustable lock-nut which is dialed to a relative depth. This nut contacts the piston housing when the piston reaches it's lowest point. This stops the polishing.
A6: I am not familiar enough with the GATAN fixture to answer this question fully. However, my experience with using a tripod type device (as Mr. Henriks suggested) with the sample mounted in the middle of the fixture produces a sample damaging instability during polishing. Shifting of the fixture from a base created by the sample and two DELRIN feet to the sample and another set of feet causes edge chipping if not carefully monitored.
If I can answer any questions for you regarding diamond lapping films, fixturing, etc., please don't hesitate to contact me directly.
Best regards,
Scott D. Holt BUEHLER, LTD. 41 Waukegan Rd. Lake Bluff, IL 60044 Phone: (847)295-4546 Fax: (847)295-7942
Here are my respones R1-6 to your comments C1-6 from my answers A1-6 to your questions Q1-6:
Response 1: As you may have read in the response from Scott Walck:
"If you use the diamond films with the GDG, when the sample becomes planar with the base of the grinder, then the grinder tends to stick to the film by surface tension because of the large area. This is even more of a problem at the lower grit sizes."
This will not happen on a soft cloth with diamond spray because you are not able to form a vacuum as you can with our film. The downside is that you will end up with rounded edges on your sample. The vacuum problem is exacerbated by using the small increments which are best to use with the gatan Disc Grinder. As you polish the sample down or use very small increments, the entire base becomes nearly co-planar which causes the vacuum to form. On the SBt fixtures, there is a space between the outside ring and the center piston which minimizes this contact area and hence the vacuum effect.
I am not surprised that Gatan recommends 3M films, they are not too likely to refer their customers to SBT! As far as the 50u increments go, that may be an advisable number to use where it minimizes rocking on the sample caused by the sample sticking out below the bottom of the fixture, but also may minimize the vacuum effect. I'm guessing that is the reason.
RE: The Tripod Polisher (R) The sof t delrin feet of the Tripod Polisher are actually made to wear out. The idea is that you don't want any material which is harder than your sample to make contact with the wheel. We use this logic in Tripod Polishing because the sample becomes very thin and fragile and we do not want any particles from WC-Co feet to containate the film. Furthermore, the Tripod Polisher (R) has the facility to adjust for the change in level of the feet. The fixtures with the tungsten carbide do not have the ability to be adjusted after polishing has begun so we try to maintain the parallelism as best we can by resisting wear. The measurements on the micrometer are, for the most part, not used. You make "relative" adjustments with the micrometers, but you do not actually measure your sample thickness with them. Sample thickness is typically determined optically.
The delrin feet on the Tripod Polisher (R) can also create a vacuum if the are polished flat and co-planar. In fact, we sometimes grind facets onto the feet to maintain more of a point contact with the film. One of our competitors sells a tripod Polisher with "non-rotating" micrometers. They charge you more for them and they are counterproductive. By utilizing non-rotating micrometers you are creating the very situation you are trying to avoid.
Response 2: If I understand this now, you are cutting your sample with 68u diamonds which would indicate a damage layer of 3 x 68 or 204u. I suppose you would need a fairly thick sample if you need to cut on both sides of the sample. I'm not sure how to avoid this. Of course, this 3x rule is a general rule and will vary with sample type. Perhaps you would like to do a little study for us in your spare time?! :} ) Assuming that you are looking for a 1u final thickness and that you will use each of these steps, the process would then be something like this:
30u: Polish until 105u thick 15u: Polish until 51u thick 6u: Polish until 21u thick 3u: Polish until 10u thick 1u: Polish until 4u thick 0.5u: Polish until 2u thick Colloidal silica for final polish.
The rough formula I use is (3 x current particle size) + particle size of next film in sequence. This gives me the amount of material I need remaining after a particular step in order to remove thedamage and still have "good" material to work with. Of course, you may want to be a little more conservative or aggressive depending on your confidence level. You can remove film steps and apply the same formula to come up with alternative sequences.
Response 3: You got me on this one. Actually, you probably got me on the other ones too, but at least I could come up with some reasonable answer for you! : {)
Response 4: No. The SBT polishing fixtures have specimen mounts that are maintain in a single orientation by way of 2 locating pins that come out of the center piston and fit into holes in the back of the removable mouting block. The piston does not rotate because there is a slot in the piston. A set screw then passes through the outside ring and into the slot to prevent rotation of the piston relative to the outside ring.
Response 5: We have a micrometer type arrangement at the top of the piston which allows you to dial in an amount of material to remove. This is done after a simple zeroing procedure. The dial at the top of the piston will stop the polishing process at the desired thickness by making physical contact with the outside ring. This prevents the piston was moving down any further.
Response 6: The Tripod Polisher is extremely effective in polishing very flat and parralel samples. You have great ability to make adjustments to correct for any errors in sample mounting and other inaccuracies in the process. However, since you have the ability to make the changes, you generally need to make them which makes the process a bit more cumbersome. You can definitely get a great sample, but you do have to work for it.
Our other fixtures rely more heavily on minimizing the variables. We have a mounting press of sorts that you use to firmly press your sample against your mounting block minimizing glue thickness and maximizing parallelism. We also recommend that you polish your mounting block while mounted in the fixture prior to mounting your sample. This ensures that your moutning block is parallel to the plane of the tungsten carbide feet. As there are no adjustments possible, it is best to follow the entire procedure carefully. Even with no adjustments, you can get a pretty darn parallel sample! Wealso make a polishing machine (Model 920) that can be fitted with workstations to hold and rotate the fixtures. We also build custom attachments (for less than you might think!) which allow a cusomer to polish 2 sides parallel to each other by simply flipping the sample over and not re-mounting.
RE: Cyanoacrylate My understanding is that any cyanoacrylate will be acetone soluble. Perhaps they just don't list acetone on the label for some reason? A sure bet is to get something like a nail bonding glue that is used to fix fingernails or attach fake nails. Check the cosmetic counter. I will try to put a tube of super glue in the mail to you, but I am not certain if it will still be good when it arrives. I have tried this before and had problems. That's the reason we stopped supplying it - it seemed a little silly to make an unhappy customer because of a $3 bottle of glue that we don't even make!
Well I think I am worn out. You have asked a lot of very good questions and it is obvious that you take your sample preparation very seriously. I am always pleased to help in any way i can and i encourage you to contact me whenever i may be of assistance. Of course, next time i wouldn't mind if talked about your application for some South bay Technology equipment! : {)
Best regards-
P.S. I would love to get a copy of other responses you get concerning this topic. Thank you!
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com sbt-at-msa.microscopy.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
I am getting ready to purchase a replacement sputter coater and I was hoping to sollicit user comments (either good or bad or general) regarding either the Pellco SC-7 Automatic sputter coater or the EMITech K550 automatic sputter coater (aka EMS 550) and/or the integration of the thickness monitors available for each unit.
PLEASE EMAIL ME DIRECTLY !!!!
ALL COMMENTS WILL BE KEPT PRIVATE !!!!
(I am not looking to publically condemn nor praise either instrument - I like both companies, but without having worked with either instrument, I'm looking for comments from those out there who have.)
Thank you in advance.
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
} We would like to know if anyone has an opinion on the effectiveness of } using a silver screen and double projectors for 3D imaging. We do not want } to commit ourselves to buying a screen (which is quite expensive) and then } find it has been a waste of money. } } We would like to hear from anyone who has tried this or any other } alternative method of displaying 3D images to groups of people.
Dear Mark, We have two large screens--they are aluminum, but look silvery; I assume these are what you want--for 3D display with polarized light & dual projectors. The images are quite spectacular and always make a big impres- sion. If you have many 3D images to display, this kind of system may be well worth the initial investment. The screens last for a very long time; ours are } ~25 years old and are as good as ever. I have no idea of the current price for these screens. Yours, Bill Tivol
We are encountering problems in disposal of uranium salts as our site safety officer considers it as a mixed/radioactive waste. I wonder what is the experience of other microscopists regarding radioactivity/disposal of uranium compounds.
Manoj MISRA Unilever Research US Edgewater NJ
Here in Bristol, we are, as long as the University Safety Office knows, and keeps some control over it, allowed to dispose of up to 100g of uranium per week (I think it is) in with the normal office/laboratory rubbish. I was surprised when I first found this out, but it has allowed us to dispose of a collection of unwanted uranium oxides and the like. Keith
I am looking for suppliers of SEM cryopreparation and cryo-transfer systems. Vendors, suppliers or colleagues with e-mail or fax numbers please respond directly to me. And yes, I have done the obligatory netsearch with very few direct scores.
Terri- A bain marie *is* a double boiler. It was invented by a medieval Jewish alchemist known as Mary the Prophetess, hence the name. Aren't you glad you asked? Steven Slap 102134,1660-at-compuserve.com
Does anyone know a good reference on the possible mechanisms of fluorescence quenching? Especially processes which involve the interaction of fluorophores with transition metal complexes or colloidal particles, from a chemistry perspective (electron transfer, molecular orbitals and so on).
I was wondering whether any of you could help giving me some tips or references where to look for counting strategy for particles (such as number of particles to be counted regarding the size and/or magnification). At this moment I'm trying to obtain polystherene particles size distributions by TEM and SEM.
Has anyone experience in working with SIGMA SCAN PRO, made by Handel, for such a distribution? and for images analysis? I would like to hear any comment about it or about any other software suggested for the subject.
Thanks a lot. Silvia Montoro CERIDE - Centre for Research and Development Santa Fe - Argentina csedax-at-arcride.edu.ar
Message-Id: {199602081547.JAA26446-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 07:39 PM 2/7/96 -0600, you wrote:
.....Do you have any info, printed or otherwise, regarding your pricing structure?
*************** No, I don't have anything formalized. I just sat down and calculated what we were spending to get one case done (costs, including salaries, supplies, overhead, etc., divided by total number of micrographs, and then how many micrographs were shot per case). To that I add what I think is reasonable to support equipment replacement. That's what I charge. For clinical cases the department adds a professional component.
I have a question on etching polysilicon on SOI material. I need to find an etch that will etch the polysilicon and not the silicon on the buried oxide. Feel free to call me or send your phone number. Thanks, Mark Windland Honeywell 612-954-2845
We followed the same procedure as Henk (using PhotoShop to produce red/green anaglyphs from pixel shifted z-sections) but then digitized the images onto slide film (Kodak Elite) using a Polaroid Palette. A single projector then did a great job (in fact it actually worked with red/blue glasses...)
Hank Colijn wrote: I have projected stereo images successfully by creating red/green anaglyphs on transparency material. We took our digital images, combined them using PhotoShop or similar program, and printed them on a dye-sublimation printer. Using a standard overhead projector and the red/green glasses, the images greatly impressed my audience. The dual projector system seems a bit specialized.
Henk
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
A few comments on this thread from our experience:
We have mounted on polarizers on the ends of cardboard tubes. The tubes are approximately the diameter of the Kodak projector lens barrel. We put a lenght of the thin packing sponge material that our hazzardous chemicals come wrapped in around the inside of the tube to provide friction and hold the tube in place. This allows the polarizers to be rotated to get maximum extinction with the glasses. We order bulk glasses for a nominal cost. Most come with the polarizers set at 45 degrees to the horizon. However, some glasses come with the polarizers horizontal and vertical. The tube system allows us to adjust or projector polarizers to match either set of glasses.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
} Date: Wed, 07 Feb 1996 16:31:59 -0500 } From: zaluzec-at-microscopy.com (Nestor J. Zaluzec) } Subject: Minor Problems } X-Sender: zaluzec-at-microscopy.com (Unverified) } To: Microscopy-at-Sparc5.Microscopy.Com } } G'day Subscribers... } } We've got a minor glitch in the subscribe/unsubscribe system. } Unfortunately, I'm on vacation and too far away to fix the link. } This just means there will be a delay if your trying to } unsubscribe. I'll be logging in remotely and doing manual } updates, but the connections are sometimes slow from here. } } I'll be back in town the middle of next week and we should } be back to normal. } } Sorry for the inconvenience } } Nestor } Your Friendly Neighborhood SysOp } } Weather report: 34 C, partly cloudy, and the view on } the beach is great. ;-) } }
Vacation? VACATION?! That's pretty cheeky. And 34C? Last week we had MINUS 34 C! Enjoy your break, Nestor.
After reading a few of the messages on this server from users in different states representing various regions of the USA, I think about the growing pains we are facing with what it seems "inconsistent regulation in the UNITED" deserves a revisit? Two items are worth recollecting: osmium and uranyl disposal. If you think that the disposal of these chemical (which by all means are hazardous) think about the follwing. During one my visits to California (NASA AMES RESEARCH CENTER) I was prohibited (and instructed so formally) from pouring SALINE (That's right PBS!) down the drain! Thus, I immediately asked where I should urinate (you know amonia, amino acids, etc). Even though the biosafety officer did not have a good answer (take that back... regulations) he was rather upset. The moral of the story: I learn quickly after arriving to the states long ago, that -the outcome of a procedure matters less than the implementation of the regulation that requires it-. That is the good news, the bad one is thatit will get worse if we do not inject a tiny bit of common sense into all of this mess and others. My two daughters were taught at school already that animals are terribly mistreated by industry and researchers, but I had to point out to her the reality behind the production of a juicy burger and/or taste fried chicken nudget? This was easy for me because when I was her age I butchered the chickens we ate and witness the killing and butchering of the pigs, cows, etc. Go on, make your contribution, and get started now-tell as really is!
********************************************************************* *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * * {Fermin-at-tmc.tulane.edu} ----} Prof. Pathology & Otolaryngology * *http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html* *********************************************************************
A visitor to our lab from Switzerland brought us an adhesive used for sealing wounds, called HISTOACRYL Blue. We have found it to be very useful and would like to find a supplier in the US. Any leads will be appreciated.
This question was asked before, and I replied direct to the poster not to the listserver. Your supplier for polyclonal nitric oxide synthetase antibody is:
AFFINITY BIOREAGENTS, INC 14818 West 6th Avenue, #13A Golden, CO 80401
We have just received the new (1996) Lindscott's Directory of Immunological and Biological reagents (Order from Lindscott's Directory, 4877 Grange Road, Santa Rosa, CA 95404; phone (707) 544-9555, Fax (415) 389-6025) which is a good place to start looking for antibodies. Some entries for anti-nitric oxide synthetase antibodies:
POLYCLONALS:
Rabbit antibody against inducible NOS, mouse macrophage:
Alexis Corp. P.O. Box 927190, San Diego, CA 92192, USA Tel: (619) 658-0065, Fax (619) 658-9224
Rabbit antibody against inducible, neuronal or endothelial NOS:
Oxford Biomedical Research P.O. Box 522, Oxford, MI 48371 Tel (in US) 800-692-4633, Fax (810) 852-4466
MONOCLONALS:
IgG2A against brain NOS, against mac, inducible NOS, against NOS EC:
Transduction Laboratories 133 Venture Crescent #5, Lexington, KY 40510, USA Tel (606) 259-1550, Fax (606) 259-1413
Against inducible NOS:
Research and Diagnostic antibodies P.O. Box 8300, Berkeley, CA 94707 Tel (510) 262-9000, Fax (510) 262-9127
Since I've never used these I can't tell you whether they work, but they are probably good places to start.
Richard D. Powell ************************************************************* NANOPROBES, Incorporated 25 East Loop Road, Suite 124, Stony Brook, NY 11790-3350, USA http://www.lihti.org/research/ecodev/incubten/nano/home.html
} } Does anyone in cyberspace know where I can obtain and antibody to Nitric } } Oxide Synthase which will work on mouse brain. } } } } Dr Terry Robertson } } Electron Microscopist } } Department of Pathology } } University of Western Australia } } Nedlands 6009 } } } } phone 346 2935 } } Fax 346 2891 } } email troberts-at-eosin.path.uwa.edu.au }
(If this appears twice, please ignore the earlier version-my mistake for E-mailing Microscopy-at-Sparc5 instead of MSA. My apologies)
Dear Christine Block:
Someone else asked about antibodies to nitric oxide synthetase, and I replied direct to the poster not to the listserver. Affinity Bioreagents supplies a polyclonal antibody to nitric oxide synthetase; their address is:
AFFINITY BIOREAGENTS, INC 14818 West 6th Avenue, #13A Golden, CO 80401
For other polyclonal and monoclonal antibodies (in reply to the earlier question):
We have just received the new (1996) Lindscott's Directory of Immunological and Biological reagents (Order from Lindscott's Directory, 4877 Grange Road, Santa Rosa, CA 95404; phone (707) 544-9555, Fax (415) 389-6025) which is a good place to start looking for antibodies. Some entries for anti-nitric oxide synthetase antibodies:
POLYCLONALS:
Rabbit antibody against inducible NOS, mouse macrophage:
Alexis Corp. P.O. Box 927190, San Diego, CA 92192, USA Tel: (619) 658-0065, Fax (619) 658-9224
Rabbit antibody against inducible, neuronal or endothelial NOS:
Oxford Biomedical Research P.O. Box 522, Oxford, MI 48371 Tel (in US) 800-692-4633, Fax (810) 852-4466
MONOCLONALS:
IgG2A against brain NOS, against mac, inducible NOS, against NOS EC:
Transduction Laboratories 133 Venture Crescent #5, Lexington, KY 40510, USA Tel (606) 259-1550, Fax (606) 259-1413
Against inducible NOS:
Research and Diagnostic antibodies P.O. Box 8300, Berkeley, CA 94707 Tel (510) 262-9000, Fax (510) 262-9127
Since I've never used these I can't tell you whether they work, but they are probably good places to start.
Richard D. Powell ************************************************************* NANOPROBES, Incorporated 25 East Loop Road, Suite 124, Stony Brook, NY 11790-3350, USA http://www.lihti.org/research/ecodev/incubten/nano/home.html
} } Does anyone in cyberspace know where I can obtain and antibody to Nitric } } Oxide Synthase which will work on mouse brain. } } } } Dr Terry Robertson } } Electron Microscopist } } Department of Pathology } } University of Western Australia } } Nedlands 6009 } } } } phone 346 2935 } } Fax 346 2891 } } email troberts-at-eosin.path.uwa.edu.au }
MR-Received: by mta GATEV3; Relayed; Thu, 08 Feb 1996 11:21:30 -0600 Alternate-recipient: prohibited Disclose-recipients: prohibited
--Boundary (ID D1sGwLyQksj+RO0n0DfJlw) Content-type: TEXT/PLAIN; CHARSET=US-ASCII
--Boundary (ID D1sGwLyQksj+RO0n0DfJlw) Content-type: MESSAGE/RFC822
The Materials Analysis group at SEMATECH in Austin, TX has an immediate opening for a TEM lab Technician. SEMATECH is a government/industry consortium working on advanced projects related to the fabrication of 0.25/0.35um CMOS integrated circuits.
For more information please contact Carolyn Gondran -at- 512-356-3149 , Philippe Maillot -at-512-356-3944 or Ted Boden 512-356-3324 or respond by direct e-mail.
JOB DESCRIPTION: Sample preparation of semiconductor materials and devices, both plan view and cross-section, for TEM. Develop improved methods of sample preparation. TEM operation to facilitate this development will be encouraged. Photographic processing and filing of TEM negatives and prints. Maintenance of sample preparation and dark room equipment and supplies
JOB REQUIREMENTS: 2+ years experience in TEM sample preparation and photographic processing required. Familiarity with tripod, dimpling and FIB techniques desired. This position requires strong organizational skills, and in particular, the ability to work on multiple tasks while maintaining precise records and tracking procedure. Experience in microelectronics industry a plus.
Associate degree in physics, chemistry or related subject.
Registered-mail-reply-requested-by: Ronald.Cohn-at-SYNTEX.COM MR-Received: by mta RVAX.MUAS; Relayed; Thu, 08 Feb 1996 17:25:49 -0800 MR-Received: by mta RVAX4; Relayed; Thu, 08 Feb 1996 17:25:49 -0800 MR-Received: by mta CHUB1; Relayed; Thu, 08 Feb 1996 17:20:30 -0800 Disclose-recipients: prohibited
List subscribers,
We need to identify apoptotic cells in cultures of cells grown in suspension. We would like to do this using the TUNEL (terminal transferase-mediated dUTP-biotin nick end labeling) technique with either fluoresence or enzyme-substrate reaction product as the read-out. I would appreciate hearing from anyone who has used this technique regarding which commercial labeling kits may be better than others, recommended protocols, etc. Thanks in advance.
Ron Cohn Structural Biology Laboratory Roche Bioscience Palo Alto, CA ronald.cohn-at-syntex.com
On March 8, 1996, a workshop on confocal microscopy will be held in the Laboratory of Physiology, KULeuven, Belgium.
Program:
08.30: Registration 09.00: Welcome: Prof. Dr. J. Janssens, Dean of the Faculty of Medicin KU Leuven 09.15: Confocal vs conventional microscopy. H. van der Voort, J. Bauman, H. Vrolijk, W. Sloos & H. Tanke, SVI Hilversum, UA & RU Leiden, The Netherlands 09.45: Spontaneously spreading calcium waves in rat cardiac myocytes: implications of the data acquisition by confocal microscopy. W. Wussling, Martin Luther University, Halle, Germany 10.15: Coffee and demonstrations on confocal microscopes 11.00: Confocal laser scanning in the reflection mode. Z. Mischal, CNRS, Villejuif, France 11.30: Free communications 12.00: Lunch 13.15: Demonstrations on confocal microscopes 14.00: Confocal fluorescence life time imaging with two-photon excitation. H. Gerritsen, J. Sytsma & J. Vroom. Universiteit Utrecht, The Netherlands 14.30: Manipulation, N-dimensional visualisation and measurement of the living cell with super-resolution and super-contrast. P. Van Oostveldt, U Gent, Belgium 15.00: Coffee and demonstrations on confocal microscopes 15.45: Free communications 17.00: Final conclusions and farewell. B. Himpens, KU Leuven
Date and location: The workshop will be held on Friday, March 8th, 1996 at the KULeuven Lab Physiology, 8th floor Onderwijs & Navorsing, Gasthuisberg Herestraat 49 B - 3000 Leuven Belgium
Contact person: B. Himpens Lab Physiology Herestraat 49 B - 3000 Leuven tel + 32 16 34 57 27 or + 32 16 34 71 46 fax + 32 16 34 59 91 E-mail: Bernard.Himpens-at-med.kuleuven.ac.be
More information and online registration: See URL: http://www.kuleuven.ac.be/kuleuven/news/wcm/
Peter Stalmans Peter.Stalmans-at-med.kuleuven.ac.be Laboratory of Physiology KULeuven Herestraat 49 B-3000 Leuven Belgium tel: +32-16-34 71 46 fax: +32-16-34 59 91
Dear All, A few of you have expressed surprise at my comment on disposal of active waste here in Bristol. I thought (though there is no direct microcsopy content - shouldn't this be in one of the safety groups??) you might like a couple of quotes from the relevant University safety handbook. "Aqueous waste. Our primary waste disposal route is via the sink... This is both the least restrictive and the least environmentally damaging" It then goes on about how this would be diluted by all the other waste water the University discharges into the sewars. "Very low levels of solid radioactive waste disposal are permitted by out authorisation from HMIP (Her Majesty's Inspectors of Pollution) to the normal waste bins.""No visible radioactive signs are to be present on the box or bag used for disposal and please try to ensure that the package does not look interesting..." There are records that have to be kept, and regulations on there being no external contamination, maximum permitted levels, only one lot per waste bin, etc.
-- Dr. Keith R. Hallam University of Bristol, Interface Analysis Centre, Oldbury House, 121, St. Michael's Hill, Bristol, BS2 8BS, England Telephone: + 44 (0)117 925 5666 | E-mail: k.r.hallam-at-bristol.ac.uk Facsimile: + 44 (0)117 925 5646 | URL: http://zeus.bris.ac.uk/~phkrh/
Message-Id: {199602091411.JAA28919-at-vaxserv} X-Sender: nnicklaus-at-cave.sarnoff.com X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Does anyone know of some "good" PC bases image processing software designed for microscopy applications? I need to do some time based studies on fluorescent DNA strands.
I have an optical trap set up to hold and maneuver the strands as attached to sub micron latex beads. I want to study the effects of various enzymes and environments on the DNA while in the trap.
Any suggestions as to software and its required (desired) hardware would be appreciated.
Message-Id: {199602091417.JAA28934-at-vaxserv} X-Sender: nnicklaus-at-cave.sarnoff.com X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Does anyone know of custom or commercially available UV microscope objectives for use in Laser Induced Fluorescence studies?
I have contacted Zeiss, Nikon, Olympus and Leica. Zeiss has the best of the group but the UltraFluars all fluoresce in my UV laser beam (266 nm). I want as high an NA as possible, i.e. immersed optics, so the reflective designs, e.g. Ealing, I have seen are not fast enough.
Fluorescence excitation wavelengths are tunable between 260 and 290 nm. Fluorescence emissions are from 300 to 500 nm.
Any suggestions would be appreciated. I can afford a custom design after I prove the concept at more limited wavelengths.
Any comments from cyberspace on the additives that EM supply houses sell to add to epoxy resins that are supposed to improve sectioning properties? I am referring to products such as Poly Cut Ease (Polysciences) or SureCut (EMS). Do people think they actually help? Are there any disadvantages?
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
With regard to using 3D aluminised screens for projecting polarised images , I can only add in support of Jim Pawley's comments on the advantages for imaging colour - in my case cell reconstructions with colour coded organelles - which come out only with the polarised system.
As regards sourcing a screen try your Chemistry colleagues who probably have one lurking somewhere as they may project molecular graphics in this way for teaching. This is where I borrow my portable screen from here in Glasgow !
Laurence Tetley Dr Laurence Tetley IBLS EM Centre Joseph Black Building University of Glasgow Glasgow G12 8QQ
I am searching for the most comprehensive articles about critical point drying techniques. All suggestions are appreciated.
Any problems with going directly from ethanol into carbon dioxide or is amyl acetate useful in between? My specimens look OK using just ethanol.
Also: I am experiencing a run of failing (most often between 400 and 800 psi) front window Dowty seals on my drying unit even if I do only 'dry' (carbon dioxide only) test runs. Is anyone else having this problem? This device has been operating for 17 years with few leaks. All sealing surfaces are smooth and clean.
Thank you in advance.
Jim Romanow Electron Microscopy Facility Physiology and Neurobiogy Department The University Of Connecticut Storrs bsgphy3-at-uconnvm.uconn.edu
I have had over a dozen requests for this information so I am posting it directly to the list:
We have always purchased our stereo glasses from Ted Pella 1-800-237-3526 outside of California They are in cardboard mounts and relatively inexpensive. However, I did not see them listed in the current catalog. I hope they still carry them. Any one know of an alternate supplier?
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
I have prototype LVSEM that employs the stage mechanism from a Philips EM 430 TEM. I would like to purchase a cold stage for this instrument if I can find one (standard Philips rod). Anyone out there about to "decommission" one?
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
Hello Everyone, I'd just like to put in my two cents on the subject of multiple users of an SEM (or in my case an electron microprobe)
Our instrument is used at least 16 hours a day and since I'm only paid for eight almost all our users are trained to use the machine independantly. This allows them faster access to the machine and saves them or their advisors money. The only people we don't encourage to become independant users are those who will only use the machine once or twice. Most users require 3-4 daytime shifts before i will let them use the machine independantly (I give them my home phone number too).
In the six years I've been running the lab I can't recall one incident where the machine has actually been damaged by inexperienced users. On several occasions it has been temporarily put out of commission (computer problems, etc) but usually I can get it back on line immediately the next day. Although there is the potential for a user to damage the instrument (i.e during a sample change) most of the rest of the instrument is fairly fool proof. It is easy to screw up your analyses, but difficult to hurt the machine.
I'm sure the machine would last longer and require less service if I was the only one operating it, but we don't have that option. I also feel that users are much better off acquiring their own data, they know what they want and can change their strategy if they find something unexpected
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Dear Bonnie, I run a Materials Engineering EM lab at the University of British Columbia, with two SEMs and a TEM. Most of the researchers who use the instruments are graduate students and Research Engineers. I have always encouraged these people to do their own research on the SEMs, as much as possible, since only they know exactly what they want and that way one me can keep three instruments optimally used. After I show them how to use the SEM, I watch and encourage them to ask me for further instruction for picture taking, higher mag, etc. Some people, who have a lot of work to do, may gain my permission to use the SEM after hours, but only after I feel they have gained a lot of experience during working hours, when I am there to supervise, and only after I have instructed them on how to properly shut down and what to do in the event of problems. I also purchased the instruments originally with ease-of-use in mind. They are fully automatic and easy to get good results on. I must feel that the person has a good understanding of the important issues and knows how to properly handle the instrument. Mind you, I don't have a field emission SEM. I have had damage, but only once in 15 years and it could have happened in normal hours.. It also helps to scare them, be "the ogre of the EM lab". Mind you, we are a teaching lab, so the learning is as important as the doing. I know many EM operators do not like the idea of letting any ham-fisted graduate student or engineer loose on their instrument, but modern SEMs really can be operated by any intelligent being. So long as you satisfy yourself as to this person's knowledge, experience and caring, I'd see no harm. Hope this helps.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Message-Id: {199602092029.OAA28451-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 10:17 AM 2/8/96 EST, you wrote:
".....to form yellow, iridescent hexagonal prisms of lead iodide....."
I stained one grid with our lead stain alone, and another with our uranyl acetate stain alone. The lead stained grid had the hexagonal deposits and the other did not. So it's not the uranyl acetate. It's interesting that lead iodide forms hexagonal crystals. *******************
".....Perhaps these are your crystals - a bit of a long shot since you did not mention iodine in our elemental analysis of the crystals...."
No, there is no iodine in either of our stains, or any of the sample preparation steps; and I did not get an iodine peak with the microprobe. *******************
I have borrowed an anti static gun like the Zerostatic Eliminator that you suggest. We are in the process of trying it out. At least the grids don't jump up and cling to the lid of the plastic petri dish any more.
These things help a great deal when using a glass knife. We use lecithin in the epoxy accelerator . ( use eual weight of lecithin and accelerator, then double the volume for use) A glass knife will last a long time if you have it in the resin. It was a great help to students just learning to section, They could get usable stuff the first day at the microtome. It can give a blotchy look to empty resin areas or those that have little density, but most tissues are "busy " enough that you never notice it. See Mollenhauer , 1986, J. EM Tech 3:217-222 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
I'd like some input, opinions etc. on the pitfalls of having multiple operators on an SEM. My situation is that I am part of a materials analysis group that provides analytical services to corporate technology. Our group consists of a number of engineers and technicians with expertise in a variety of analytical areas. Our company has recently hired a young PhD to do materials research in another department . He is pushing very hard to be allowed to use our SEM at night and on weekends. He has 'used an SEM before' and doesn't see a problem in using ours. It has always been our practice to encourage researchers to come into the lab while their samples are being analyzed, but not to allow individuals to use the instrument themselves. I believe that the analysis is done more effectively by individuals whose expertise is in that area. However, management appears to be leaning towards changing the rules for this individual, and having me train him on the instrument.
We have a four year old JEOL 6400 instrument with a Link eXL analyzer, UTW detector, complete stage automation. We also have a computer system attached for collecting digital images, we do not use polaroid film. While this individual has used and SEM he has not used either a JEOL or LINK system before. I feel that training would be extensive. This is currently the only SEM we have. It is used approximately 12 hours per day by myself and my staff. Quite often EDS analysis is carried out using computer control and stage automation overnight. However, management feels that if someone is willing to utilize the instrument in those few hours a week when it is not being used, they need to take advantage of that to receive a maximum benefit from the asset.
Several years ago we had two older SEMS and due to 'corporate downsizing', we had only one operator. At them time I trained approximately 2 dozen engineers who desired to use the 'extra' SEM on their own. We disposed of the instrument about a year ago when it had not been touched for almost a year. The time I spent in training all of those individuals was probably greater than the time they as a group spent using the instrument. Hindsight being perfect I wish we had kept the old instrument!
Anyway, I would appreciate any input both pro and con from the list regarding opening up an instrument to multiple infrequent users.
For your information, the following two companies also sell the “wedge-polishing” tool, supplies, and various consumables (diamond lapping films/suspensions, mounting wax/glues, etc.) for TEM specimen preparation of materials:
UltraMetrix 4802 E. Ray Rd., No. 23-267 Phoenix, AZ 85044 (602) 706-5745 FAX (602) 496-6505 e-Mail ULTMTRX-at-aol.com
Allied High-Tech Products P.O. Box 4608 2376 E. Pacifica Place Rancho Dominguez, CA 90220 (310) 635-2466 FAX (310) 762-6808
Rene E. Nicholas Spec. Prep. Workshop Coordinator AMC Group
At 1:09 PM 2/9/96 -0500, Bonnie Davis - Kennametal Inc. wrote: } I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM. My situation is that I am part of a materials } analysis group that provides analytical services to corporate } technology. Our group consists of a number of engineers and technicians } with expertise in a variety of analytical areas. Our company has } recently hired a young PhD to do materials research in another department . } He is pushing very hard to be allowed to use our SEM at night and on weekends. } He has 'used } an SEM before' and doesn't see a problem in using ours. It has always } been our practice to encourage researchers to come into the lab while } their samples are being analyzed, but not to allow individuals to use the } instrument themselves. I believe that the analysis is done more effectively } by individuals whose expertise is in that area. However, management } appears to be leaning towards changing the rules for this individual, and } having me train him on the instrument. } } We have a four year old JEOL 6400 instrument with a Link eXL analyzer, } UTW detector, complete stage automation. We also have a computer system } attached for collecting digital images, we do not use polaroid film. } While this individual has used and SEM he has not used either a JEOL or } LINK system before. I feel that training would be extensive. This is } currently the only SEM we have. It } is used approximately 12 hours per day by myself and my staff. Quite } often EDS analysis is carried out using computer control and stage } automation overnight. However, management feels that if someone is } willing to utilize the instrument in those few hours a week when it is } not being used, they need to take advantage of that to receive a maximum } benefit from the asset. } } Several years ago we had two older SEMS and due to 'corporate } downsizing', we had only one operator. At them time I trained } approximately 2 dozen engineers who desired to use the 'extra' SEM on } their own. We disposed of the instrument about a year ago when it had } not been touched for almost a year. The time I spent in training } all of those individuals was probably greater than the time they as } a group spent using the instrument. Hindsight being perfect I wish we had } kept the old instrument! } } Anyway, I would appreciate any input both pro and con from the list } regarding opening up an instrument to multiple infrequent users.
Bonnie, Since we encourage use of the JEOL TEM and SEM by trained users, we run into this situation often. Currently, we require training on our instruments plus a minimum of 15 hours of daytime use prior to an approval of use during off hours. The principal investigator (of the grad. student or postdoc) is also asked to agree to cover costs should any malfunction necessitate lengthy (costly) alignment procedures. I constructed a form for this purpose and the signed agreement is kept with the log book at the scope. This has given me some control over the situation and those users who agree to our demands are usually competent users. If the SEM uses a LaB6 or is a FE, I would be more cautious, i.e., demand more daytime use and obtain written consent to replace the crystal/filament. One last thing to consider---an estimate of costs incurred by your unit during the required "daytime" use if your unit does not charge other units for instrument use. Regards, Rosemary Walsh
Hello all, I am currently doing TEM of ferroelectric materials (PZT, PST etc.) and seem to be seeing domains or domain walls in thin TEM samples but not in thick ones. I know that TEM of ferroelectrics has been done for some years - does anyone on the listserver know of any references which describe the contrast mechanism which makes them visible in TEM?
Many thanks in advance,
RIchard Beanland, GMMTL Caswell, Towcester, Northants NN12 8EQ, UK Tel. +44 1327 356363 Fax. +44 1327 356775 Email richard.beanland-at-gecm.com
All modern EDS systems include in the quantitative procedures an option "standardless analysis". When standardless analysis is performed, tipically the analysis of stainless steel is demonstrated with measuring transition elements Fe, Cr, Ni, Mn, Co and V. The K-lines of these elements span in the range 5 - 7.5 keV where the spectrometer efficiency is constant and close to unity. The product is well behaved, and a reasonably accurate analysis can be obtained, especially if the procedure has been "optimized" for steel analyses. For testing standardless programs we need suitable data base of measured intensity data with detailed information about analysis (accelerating voltage, geometry) and detector constants. Unfortunately no systematic collection of such data to be available in the area standardless analysis. This is a sad situation in comparison with , for example analysis with standards were there is an large collection of data. For this reason we are in our laboratory decided to collect the intensity data for testing standardless programs. Your help in adding new data sets would be much appreciated. The rules for the data in the collection are very simple:
a) only experimental data from certified samples are included.
b) all data must be collected under known experimental conditions: acclerating voltage, geometry (tilt angle, take-off angle) and detector constants (Be window or other thickness, Au layer thickness, Si dead layer thickness and Si crystal thickness).
c) only pure intensity data (after background substraction and peak deconvolution procedure) are included.
d) weight fraction of analyzed elements must be known.
Final database will be available at EMMPDL (Electron Microscopy and Microanalysis Public Domain Library) on anonymous FTP server WWW.AMC.ANL.GOV (Argonne National Laboratory, IL, USA). We have no any commercial interest with this database.
Thank you.
Henrik Kaker Metal d.o.o. SEM/EDS Laboratory Koroska c.14 62390 Ravne Slovenia
Hello all, I am currently doing TEM of ferroelectric materials (PZT, PST etc.) and seem to be seeing domains or domain walls in thin TEM samples but not in thick ones. I know that TEM of ferroelectrics has been done for some years - does anyone on the listserver know of any references which describe the contrast mechanism which makes them visible in TEM?
Many thanks in advance,
RIchard Beanland, GMMTL Caswell, Towcester, Northants NN12 8EQ, UK Tel. +44 1327 356363 Fax. +44 1327 356775 Email richard.beanland-at-gecm.com
IS the Gatan Ion mill still available? Joe Geller 508 887-7000
On Thu, 11 Jan 1996, Gordon L. Nord Jr. wrote:
} Dear Microscopists, } } Because of the recent downsizing of the US Geological Survey the TEM lab } must go. In addition, because the last remanents of the Bureau of Mines (may } it rest in peace) are coming to the USGS in Reston the lab space is needed for } other purposes by next week (gasp). All or part of the following are available } for transfer to US academic institutions. The USGS has not mentioned any } possibility of help with respect to shipping but I could be surprised. } } (1) JEOL 200B Transmission Electron Microscope in working condition (1974) with } STEM attachment (1978). } } (2) Multiple stages including Be double-tilt stage by Gatan and heating stage } by JEOL. } } (3) Technics Ion Mill, 1974 (works) } } (4) Gatan Duo Ion Mill with low voltage guns and cooling stage, 1983 (works) } } (5) Tracor Northern 2000 Multichannel analyser and detector 1978 (works) } } (6) LogEtronics enlarger 1974 (works) } } Contact me at the following address. } } Cheers, } Gordon } } } Gordon L. Nord Jr. } 959 National Center } U. S. Geological Survey } Reston, VA 22092 } } Office: 703-648-6745 } FAX: 703-648-6789 } } gnord-at-mactem.er.usgs.gov } }
Dear Microscopists Could anyone advise me on current developments in the embedding (wax or otherwise) sectioning and staining of botanical specimens for histological and anatomical examination. We have got as far as Histoclear, but most other techniques seem to be more than fifty years old [and working satisfactorily, I may add]. Some of the old stains are fading though and we have to replace old slides, and this might be an opportunity to apply any up to date technique.
Cheers
Stephan Helfer Royal Botanic Garden Edinburgh Scotland UK
I must echo Bob Craig's observations about "external" users. He could have easily written that word-for-word as a fellow employee here. Our back-up plan for extended use of high-demand instruments is to go to an informal swing schedule. Fortunately we have not had to resort to that officially, but most of our gang have a lot of "after hours" time logged. One method we have utilized for increasing throughput on our "automated" instruments is to dial in from home with Timbuktu and AppleTalk Remote Access to check on progress. If something has gone amuck, we can either fix it remotely or zip in to work, fix it and resume the experiment. It's not pleasant to come in at midnight instead of going to bed, but our customer's anxiety (hence ours!) is greatly diminished the next day.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Subject: Time:11:04 AM OFFICE MEMO Antibody to TRH Date:2/12/96
Hello Everybody, I am looking for antibody to Thyrotropin-releasing-hormone for immunocytochemistry which will work on rat brain. Does anyone know where I can get it? I. Polyakov NASA-Ames Research Center FAX: (415)604-0046 Email:Igor_Polyakov-at-qmgate.arc.nasa.gov
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
This has been a problem for us at times. The real issues are; the ability of people to work together, and the cost of maintenance, and keeping the instrument in top condition. We find it possible to have up to about 3 operators, as long as they can work together well. They also need to be in the same department (i.e. financed together). Pride in the condition of the instrument and the quality of results is critical.
There can be other issues, like job security. If allowing someone access you are putting someone else out of work...........
When the machine breaks, whoever had their hand on it, must immediately get it fixed. This works best when there's a service contract.If there's an assigned technician, this may not be a problem for you.
My overall impression is that the number of operators must be limited to a couple who can work together well, and not quibble over who did what.
{ } { { } } ===========} Dave King { { { } } } ------------------------------------------------------ { { { {
We have a researcher who wishes to determine if coal fly ash is present in river water and river sediment. By LM I detect some "pearly" particulates not present in control river sediment. Next step will be examination by SEM.
My question: is there an established protocol for detecting and quantifying fly ash in river water and sediment?
I will post any responses not sent to the server directly - unless otherwise directed. Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Message-ID: {199602122331.SAA16597-at-IndyNet.indy.net} To: Richard Beanland +44 1327 356363 {richard.beanland-at-gecm.com} , Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
[various snips] } I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM. } Our company has } recently hired a young PhD to do materials research in another department . } He is pushing very hard to be allowed to use our SEM at night and on weekends.
He has to be trained very well to use the machine when no staff mem- ber is around. It is much easier to have him use the SEM during the day, and I would insist that he do so after training until you are absolutely satis- fied that he is competant before allowing him to work without a net.
} He has 'used } an SEM before' and doesn't see a problem in using ours. It has always } been our practice to encourage researchers to come into the lab while } their samples are being analyzed, but not to allow individuals to use the } instrument themselves.
For our facility--a NIH biotech resource--we have several clas- sifications for outside users, from novice, for whom the staff does every- thing, to expert, who can use the instrument without supervision. We train the user in many steps, the first being the basics for tilting and transla- ting the specimen, focussing and taking a picture, then changing specimens, later changing film, starting and shutting down the scope, etc. All this takes several months for an in-house user, and longer for someone who visits occasionally.
} This is currently the only SEM we have. It } is used approximately 12 hours per day by myself and my staff. Quite } often EDS analysis is carried out using computer control and stage } automation overnight. However, management feels that if someone is } willing to utilize the instrument in those few hours a week when it is } not being used, they need to take advantage of that to receive a maximum } benefit from the asset.
There is a large negative benefit if anything goes wrong; try to impress management that a conservative approach is best.
} Anyway, I would appreciate any input both pro and con from the list } regarding opening up an instrument to multiple infrequent users.
This will take a *lot* of your time. Good luck, you'll need it. Yours, Bill Tivol
Yes, there are various programs on the market. Depending on your budget, and your need for continued support. Probably the most ecconomic of the bunch is Image Pro Plus, soon to be available using 32 bit, and windows 95.
The second PC based system, that I find very intuitive and has excellent support is Kontron's KS 400 IA system. Kontron KS versions are distributed and supported by Zeiss, Thornwood NY. The best person to contact is Uli Kolhaaus, or Elise Shumpsy at Zeiss.
Gregory Argentieri Sandoz Pharmaceuticals Corp East Hanover, NJ 201-503-8617
Hello everyone, I have been embedding pinon pine root tips for mycorrhiza research. We are close to solving most of our problems but still seem to have some left. And I was wondering if some fresh ideas may help. Problem: Portions (it seemed to be mainly the cortical tissue) of the root tip will not completely infiltrate and polymerize. The mantel looks okey and central vascular area seems to embed okey. The cortical cells seem to be our main source of infiltration problems. Some of the samples float (even after hours or days in a vacuum). At first, we used Spurrs resin with graded Etoh. We have not used acetone or propylene oxide (yet).
Figuring we were not getting all the water out of the tissue nor getting our resin into the tissue we have tried the following procedures: 1. Cut our root tip sample even smaller. 2. We obtained some Z-6040 which has been mentioned several times on this list for helping to infiltrate impervious biological specimens. We used it at 1% as advised by Virginia Lindley's article. But we also continued its use in the LR White resin as recommended by Stacie Kirsch at EMS. 3. We found we had to extend our Abs. ETOH times to over night and several days of changes of LR White. We switched to LR White as recommended in Lindley's article.
At this point, we had some success, but still had some root tips that wanted to float (even under vacuum). I have not worked with root tips or mycorrhiza before, but I may be doing more in the future. Do we need to use acetone or propylene oxide to remove all the water?
Thank-you for any comments or helpful suggestions. If you should like respond to me at my local address, I shall summarize for the listserver.
Good day, Marilee
Marilee Sellers Manager, Electron Microscope Facility Northern Arizona University Flagstaff, AZ 86011 E-mail marilee.sellers-at-nau.edu
I am searching for the most comprehensive articles about critical point drying techniques. All suggestions are appreciated.
Any problems with going directly from ethanol into carbon dioxide or is amyl acetate useful in between? My specimens look OK using just ethanol.
Also: I am experiencing a run of failing (most often between 400 and 800 psi) front window Dowty seals on my drying unit even if I do only 'dry' (carbon dioxide only) test runs. Is anyone else having this problem? This device has been operating for 17 years with few leaks. All sealing surfaces are smooth and clean.
Thank you in advance.
Jim Romanow Electron Microscopy Facility Physiology and Neurobiogy Department The University Of Connecticut Storrs bsgphy3-at-uconnvm.uconn.edu
Message-Id: {199602122054.OAA28915-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Thanks, Gib, for your message.
"....that some stray electrostatic charges floating around on these low humidity winter days can precipitate beautiful hex lead crystals just doesn't click for me...."
As I had said before, I also don't think that static charges would cause inappropriate stain precipitation, but then as you can see from the response on the listserver, a lot of people have experienced this problem and the cause(es) have not been determined. At this point I am willing to try anything and the suggestion didn't seem unreasonable. ***********************
"....I say chuck the whole damn works and switch to the Ted Pella "Grid Stick" kit to do your uranium and lead staining...."
We have tried Pella's Grid Stick. However that procedure washes the sections off of our grids. Our type of work necessitates the use of large sections and 200 mesh "Super Grids" (grids with very thin grid bars). The use of these grids results in much reduced area of contact between grid and section, and the sections just do not adhere to the grids well. Now, if you have any suggestions as to how we can get the sections to stick to the grids better, I would appreciate them very much. Then perhaps we could institute a more vigerous wash. ***********************
"....For lead, I use the Sato triple lead stain, not straight Reynold's lead citrate...."
I do know of the Sato tripple lead stain, but would appreciate the reference, if you have it handy. We have tried both the Reynold's recipe as well as the recipe from Dr. Chandler Fulton. I don't have that reference. However nothing could be simpler. To 10 ml of decarbonated, distilled water you just add 0.02 gm lead citrate powder (EMS cat no. 17800), followed by 0.1 ml of 10N NaOH. This stain worked very well for me throughout my graduate school experience.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
I'd like some input, opinions etc. on the pitfalls of having multiple operators on an SEM. My situation is that I am part of a materials analysis group that provides analytical services to corporate technology. Our group consists of a number of engineers and technicians with expertise in a variety of analytical areas. Our company has recently hired a young PhD to do materials research in another department . He is pushing very hard to be allowed to use our SEM at night and on weekends. He has 'used an SEM before' and doesn't see a problem in using ours. It has always been our practice to encourage researchers to come into the lab while their samples are being analyzed, but not to allow individuals to use the instrument themselves. I believe that the analysis is done more effectively by individuals whose expertise is in that area. However, management appears to be leaning towards changing the rules for this individual, and having me train him on the instrument.
We have a four year old JEOL 6400 instrument with a Link eXL analyzer, UTW detector, complete stage automation. We also have a computer system attached for collecting digital images, we do not use polaroid film. While this individual has used and SEM he has not used either a JEOL or LINK system before. I feel that training would be extensive. This is currently the only SEM we have. It is used approximately 12 hours per day by myself and my staff. Quite often EDS analysis is carried out using computer control and stage automation overnight. However, management feels that if someone is willing to utilize the instrument in those few hours a week when it is not being used, they need to take advantage of that to receive a maximum benefit from the asset.
Several years ago we had two older SEMS and due to 'corporate downsizing', we had only one operator. At them time I trained approximately 2 dozen engineers who desired to use the 'extra' SEM on their own. We disposed of the instrument about a year ago when it had not been touched for almost a year. The time I spent in training all of those individuals was probably greater than the time they as a group spent using the instrument. Hindsight being perfect I wish we had kept the old instrument!
Anyway, I would appreciate any input both pro and con from the list regarding opening up an instrument to multiple infrequent users.
In a commercial organization, an instrument is a resource for all. The question can not be is a person allowed to use a resource because they are or are not a member of a particular work group. The question must be how do we maximize productivity. Working within this framework, here are two observations to balance:
It is typically counter-productive to have relatively inexperienced users operate instruments which are either finicky or complex.
It is typically counter-productive to have expert users repeatedly perform a relatively simple analysis.
Now for the reduction to practice. We have trained researchers from a variety of groups to use our microscopes, OM, SEM, and TEM. This has included training an inexperienced researcher on a FEG-SEM. We have never had a mishap which cost us more than a few hours of instrument time. The OM and SEM training has benefited productivity. However, TEM training has been another story. Inexperienced researchers were repeatedly retrained, resulting in an inefficient use of expert personnel.
Good luck to all,
Craig Garrison The Dow Chemical Company Bldg. B-1225 2301 N. Brazosport Blvd. Freeport, TX 77541
I am looking for a postdoctoral researcher to fill the following post. Please could any interested parties contact me directly.
Thankyou, Amanda Petford-Long.
************************************************************* Advertisement for postdoctoral research post in the Department of Materials, University of Oxford, UK funded by Hewlett-Packard.
A post is available, in the first instance for 1 year, to study layered films for magnetoresistive read head applications. This will form part of an ongoing collaboration with Hewlett-Packard Laboratories in Palo Alto, involving both experimentalists and modellers within the Department of Materials.
The research will involve the characterisation of the microstructure and magnetic domain structure, by electron optical techniques, of films grown at HPL by sputter deposition. The films under investigation will be spin-valve structures, and the interest lies in the role of microstructure on giant magnetoresistance and magnetic properties. The microstructure of the films will be analysed using high resolution electron microscopy, and we have already had some considerable success in quantifying the nature of the interfaces between the layers in the spin valve using imaging processing techniques on HREM images, and would hope that this work could be continued. The magnetic domain structure of the films will be studied using our extensive Lorentz transmission electron microscopy facilities. These include facilities for in situ magnetising and heating, and also for the collection of quantitat- ive magnetisation maps. Some experience in electron microscopy (preferably HREM) is therefore essential.
The post is fully funded by HPL on a rolling grant basis, so that further funding after the 1 year period is extremely likely but cannot be guaranteed at this time.
The post is available from 1st July 1996 and will be on the University salary scale GBP14,317 - 21,511 p.a.
Interested applicants should submit their applications, including a curriculum vitae and the names and addresses of two referees to the Administrator, Department of Materials, Parks Road, Oxford OX1 3PH, UK. Please quote ref. AKPL. The University is an Equal Opportunities employer.
For informal queries please contact Dr Amanda Petford-Long in the Department of Materials. e-mail address: amanda.petford-long-at-materials.ox.ac.uk
Others you might try:Arnel Products Company (Tel. 212-620-4622), Biogenesis Ltd. (Tel. 603-887-4600, Email biogenesis-at-ltd.co.uk), Chemicon International (Tel. 800-437-7500), Eurodiagnostics Lab (Sweden, fax 46-409231000), Cappel-Organon Technica (Tel. 800-523-7620) and UCB Bioproducts (carried by Accurate Chemical and Scientific Corp, see above).
All these were raised in rabbit, so they should work on rat brain, although I havn't tried any.
This is from the 1996 Lindscott's Directory of Immunological and Biological reagents (Order from Lindscott's Directory, 4877 Grange Road, Santa Rosa, CA 95404; phone (707) 544-9555, Fax (415) 389-6025), one of the best places to start looking for this kind of information.
Richard D. Powell ************************************************************* NANOPROBES, Incorporated 25 East Loop Road, Suite 124, Stony Brook, NY 11790-3350, USA http://www.lihti.org/research/ecodev/incubten/nano/home.html
Message-Id: {199602131409.KAA17548-at-Snoopy.UCIS.Dal.Ca} Comments: Authenticated sender is {RMACKAY-at-ac.dal.ca}
RE: Multiple Users
Dear Bonnie,
We have operated a JEOL 733 electron microprobe ( presently with a Link eXL system ) for the past 11 years, and prior to that a Mark V. Our policy has always been to allow multiple users including graduate and undergraduate students. In 25 years we have had only one bad experience.
Best Regards,
Bob MacKay Robert MacKay Department of Earth Sciences Dalhousie University Halifax, Nova Scotia, Canada B3H 3J5 Tel: 902 494-7087 e-mail rmackay-at-ac.dal.ca
} Marilee, A microwave protocol is especially well suited for your work. I } recommend checking the recent article by Giberson RT, Demaree RS, Jr. Microwave } fixation: understanding the variables to achieve rapid reproducible results. } Microsc Res Tech 1995;32: 246-54. } } Gary R. Login, Beth Israel Hospital, Pathology } 617-667-2034; glogin-at-bih.harvard.edu
Gary, This is one alternative I hadn't even considered. Thank-you. I do not have a research microwave. But will see if ours would do. Marilee
Marilee Sellers Manager, Electron Microscope Facility Northern Arizona University Flagstaff, AZ 86011 E-mail marilee.sellers-at-nau.edu
I have a question which maybe someone can answer. Assuming constant angle during ion-beam thinning, which of these two leads to less nett damage (point defects etc) within a sample, assuming that both remove the same amount of material: a) A longer time at low energy b) A shorter time at higher energy ?
You can also try Visilog from Noesis Vision Inc. It provides the widest selection of algorithms on the market along with an easy to use GUI and macro language. We are soon coming out with a 32 bit version on both Unix and Windows NT. You can contact me for more information. Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.cam.org/~noesis 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada
} Marilee, } } A year or so ago, we were working on a similar sounding project looking } at mycorrhizal associations in pine roots (different pine). We were } interested mainly in the composition of some inclusions but prepared material } for general morphology too. We used a "standard glut/Fa" primary fix and an Os } tet secondary followed by acetone dehydration and embedment in Spurr'Us or } our own variant thereof (VCD and Quetol). We had no problem with infiltration } or sectioning. } } Here's my twenty questions: } } 1. Do you use vacuum during fixation? I generally bounce the tissue } during primary fixation by cycling the vacuum until the tissue stays } submerged up to the boiling pressure of the fixative.
We have not used vacuum during fixation. Most of the time the tissue did not float until after the Abs. Etoh. We shall include this.
} 2. Did you try to embed any of the floating material? I generally don't } try to embed "floaters". My thinking is that if they have enough air in them } to float after Os tet density increase, they've got too much air for good } transfer of any thing into or out of the tissue.
We did embed some floaters. And they were poorly infiltrated.
} 3. When do you notice the problem in infiltration? i.e. during } sectioning, or after observation. I've had occasional problems with LR white } bonding to cell walls. Usually we can get enough to hold still for a picture } if we carbon coat them after labeling and staining and before viewing. The } stuff is supposed to be able to handle some water as an impurity, but I'm not } sure I believe it. } 4. What kind of knife do you use? I used glass since our root samples came } from the real world and had entrained some sand as well as air. If yours don't } have anything hard in them (or you'Uve got lots of money;-)) might use a } diamond if youUre ot now. I cut a lot of hydroponically grown roots for class } and generally get O.K. sections in regular resin with my diamond knife. Even } they hydroponically grown tips have considerable exhaustible air in them.
We notice the problems immediately upon sectioning. These root tips are from sandy soil, and the tips are cleaned as much as possible before processing. As a precaution, we use glass knives to cut them.
} John Heckman } TEM supervisor } Center for Electron Optics } Michigan State UniversityMarilee, } } heckman-at-pilot.msu.edu
Thank-you John. We will be incorporating your suggestions. Marilee Marilee Sellers Manager, Electron Microscope Facility Northern Arizona University Flagstaff, AZ 86011 E-mail marilee.sellers-at-nau.edu
View From A University Central Service Microscopy Lab
As the Director (and only employee) in a lab with a TEM, SEM, confocal, imaging equipment, photographic darkroom, and the usual ancillary equipment, and no service contracts, I could not function if I had to do everything for users. Ninety percent plus of my users operate the equipment themselves. However, less than 5 users are 100% qualified to operate the EM's when I am not present. I insert and remove specimens, and for the TEM, process the negatives. Obviously, users undergo training, and during training one can pick up cues as to the user's future behavior. Our equipment is very user friendly, and that makes a big difference. Obviously in industry where down time is unacceptable, this approach would not work. Bruce Cutler, Microscopy Lab, Univ. Kansas, Lawrence
We have a couple of users who want to look at calcium and/or pH changes on suspension cells (culture cells). Does anyone know of specific tricks for trying to stabilize single/suspension cells in a perfusion chamber. We would like to follow a single/small group of cells throughout treatment with different agents. We have tried both charged and coated coverslips (silane and gelatin) without sucess. Any and all help would be greatly appreciated.
TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
} The silicon 'dead layer' in a detector is a mathematical concept to } describe a region in the detector of reduced sensitivity to the incoming } x-rays (the Glasgow people coined a term for this which now eludes me). } The 'dead layer' varies with the detector bias, and cannot be measured } directly - it is possible only to deduce it from experiments fitting } measured bremsstrahlung shapes to theoretical predictions.
Dear Tony, The dead layer of a Si detector can, indeed, be measured experi- mentally--at least in principle. Using an alpha source, measure the energies when the alphas are incident on the detector both normal to its face and at another angle for which the alphas still penetrate the dead layer. From these measurements the thickness of the dead layer (plus the gold) can be determined--at a number of bias voltages if desired. Personally, I'd never subject my detector to this measure- ment, but it is possible. We used to do this all the time for detec- tors when I was in grad school. Yours, Bill Tivol
This is an interesting problem. I have had to train other users to use my SEM, but have always found it to be a waste, since they didn't use it enough to remember the training. I had to go over things each time they wanted to use it.
However, if you are forced to let this person use the SEM, here are some guidelines. Mark the saturation point clearly, and make it a rule to not go past that mark. Have a log book handy for the part time user to record any 'unusual' occurences in operation, or to write questions down. Have an emergency number to call for help. (I know this is a pain for you, but could save on instrument downtime and repair costs.)
That's all I could think of right now, but I hope it helps.
Regards, Melanie Behrens Senior Chemist Texaco, Inc. Beacon, NY behrema -at- Texaco.com
Hello all, I need to obtain the address and fax number of 'Triarch Slides' in Wisconsin, USA if they are still in business. Any help would be appreciated. Regards,
Via: uk.ac.bbsrc; Wed, 14 Feb 1996 10:28:03 +0000 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 14 Feb 1996 10:29:49 +0000 X400-Received: by mta arcs.bbsrc.ac.uk in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 14 Feb 1996 10:29:49 +0000 X400-Received: by mta LARS.MUAS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 14 Feb 1996 10:27:28 +0000 X400-Received: by mta LARS in /PRMD=UK.AC/ADMD= /C=GB/; Relayed; Wed, 14 Feb 1996 10:27:31 +0000 X400-Received: by /PRMD=UK.AC/ADMD= /C=GB/; converted (ia5-text); Relayed; Wed, 14 Feb 1996 10:27:45 +0000 Notification Requested) (IPM Return Requested)
Dear All,
Many thanks to all those who responded to my query re mode of action of toluidine blue in quenching autofluorescence, particularly Bill Tivol and Bridget Southwell who gave an answer that I can trot out if ever asked again! In response to those who requested further details of the way I use tol. blue: it is applied as a 0.01% (w/v) solution made up in phosphate-buffered saline (pH nominally 7.3 - 7.6) to the sections (I routinely use 6 um thick methacrylate-embedded Aesculus hippocastanum root sections). Leave the stain to impart a 'light-blue' colour to the sections, ie, let them 'just stain', that takes about 90 sec with my tissue. Sections then mounted in anti-fade mountant and viewed - stored if necessary in a fridge (at c. 4 C). [Sections having been processed for visualisation of tubulin using FITC-linked secondary antibody, and usually stained with DAPI prior to tol. blue-quenching] "It works for me"... I do not know if tol. blue itself fluoresces under some excitation wavelengths, but it does reduce some of the background/native autofluorescence present in my tissue (for Sally Stowe). The reference I have used for this procedure is Baluska, F, Parker, JS, Barlow, PW, 1992. Specific patterns of cortical and endoplasmic microtubules associated with cell growth and tissue differentiation in roots of maize (Zea mays L.). J. Cell Sci. 103, 191-200. Of course, if anybody can provide an earlier (more definitive?) reference to its first use, I would be grateful. Hope the above is useful,
I'm posting this request for a friend that needs to buy a light microscope. He specifically is looking for a microscope with a large stage that will allow a long working distance for using a micromanipulator. Does anyone have a used microscope that they want to sell? Also, does anyone know the address or telephone number of equipment brokers that might have light microscopes? If so, please respond to Irv Widders, Michigan State University, E-mail 22457iew-at-msu.edu
I have found that (a), a longer time at low energy, is better for my samples- III-V semiconductors.
Ciara
On Tue, 13 Feb 1996, L.D.Marks wrote:
} I have a question which maybe someone can answer. Assuming } constant angle during ion-beam thinning, which of these two leads to } less nett damage (point defects etc) within a sample, assuming that } both remove the same amount of material: } a) A longer time at low energy } b) A shorter time at higher energy } ? }
Hi Bonnie, One point that everyone seems to have missed on this subject of multiple users is output quality. There is no substitute for the skill gained with experience. We tried running two SEM's in our lab, and found the overall quality of the images produced by our "trained" multiple users was not as good as the images produced by an experienced technician.
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
There has been alot of comments on the matter of multi-user of and SEM. I have been on both sides of that fence. I operated and SEM for years at the Bureau of Mines until I took a research position. With my position I had the need for SEM work. This I was able to do most by myself. I am now with a company that does not have an SEM so I have to use the one at the university. While I was the operator I was very choosey about who could use it. Only a hand full of people asked to use it and when I started the training they decided it was to much to remember and that I should do it because they did not have the time to do it correctly. Now that I use someoneelses I am very carful to follow all the instructions, I want to be able to come back. I also felt very uncomfortable the first few times I was left alone with this instrument, now I am familiar with it and use it much more. I guess what I am trying to say is some people have no idea about an instrument and are hot shot scientist, newly graduated PhD's and others. These are people to be aware of. Multi-users are okay depending on the type of business you are in and the personnal. I thought an opion from the other side should be expressesd.
Clarissa Vierrether Analytical Development Chemist The Doe Run Company P.O. Box 500 Viburnum, MO 65566 1-573-244-8109
As Tony Garratt-Reed pointed out the 'dead layer' in an EDS detector is an unfortunately chosen name. As first shown by Goulding (Nucl.Instrum. and Methods 142, 213, 1977) the 'dead layer' is simply the region over which drift anf diffusion approximately cancel each other. The behavior of this region is crucial when good resolution at low energy ( {2keV) is required. The MAS Fiori Memorial volume "Xray Microanalysis in Electron Beam Instruments" published by PLENUM contains two papers which discuss this in detail, and also contain a bibliography of relevant references - one
Joy DC, "Modeling the Energy Dispersive X-ray detector", in "Xray Spectrometry in Electron Beam Instruments", ed D B Williams, J I Goldstein and D E Newbury, (Plenum Press:New York), 53-64, (1995)
which discusses a Monte Carlo simulation of this type of effect, and a second by Jon McCarthy of NORAN which examines the practical implications of 'dead layer' behavior. The dead layer can be estimated in several quite convenient ways, including spectral measurements, matching to simulation ot, best of all, by an electron beam measurement described in
Joy D C, "The EDS Detector - A Quantitative Model", Rev. Sci. Instrum. 56, p1772-1779, 1985
As an interesting aside note that the dead layer in silicon at room temperature is only 700 angstroms or so compared to 2800 A or more at liquid nitrogen temperatures
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Bonnie,
The Integrated Microscopy Resource (IMR) is an NIH biomedical research technology resource. We have a Hitachi S-900 FESEM with cryo-capability and a YAG backscatter detector. As one of the few microscopy facilities open for biologists, the novel instruments at the IMR are made available to users world wide for biomedical research projects. Our user base consists of senior investigators, post-docs, and graduate students.
As an NIH resource, the use of instruments is reserved for medical research projects that require the specialized aspects of the available instruments. Our SEM usage policy consists of three levels of approval: basic use, backscatter detector use, and cryo-stage use. I provide user training and supervise the use of the SEM for the first 20 hours. After this time, approved users may be upgraded to unsupervised users who are permitted to work independently after hours. Facility keys (either short or long term) are available for unsupervised users. For this system to work it is critical to give each user comprehensive instructions on use of the equipment, to establish clear rules with regard to general use of the facility, and most importantly to have an individual with the authority to monitor and enforce these rules.
Our open use policy has been in place for 9 years with the SEM and 25 years at our facility. Project approval and user training prior to equipment usage has resulted in much success and very few problems. When problems have arisen the solution has been for me to re-train the user and continue to supervise them for longer time.
Hope this information answers your question.
Ya Chen
========================================================================= \ / Assistant Researcher, Cryo/SEM Coordinator TEL : 608-263-8481 \ / __ Integrated Microscopy Resource (IMR) FAX : 608-265-4076 \/ / / University of Wisconsin-Madison / / / 1675 Observatory Drive #167 Email:YChen-at-macc.wisc.edu / /__/_ Madison, WI 53706 Email:chen-at-calshp.cals.wisc.edu
IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr/imr.html IMR Symposium on integrated microscopy '96: Sept. 20-22, 1996 =========================================================================
Anyone have any good references re: Calicivirus? Especially interested in classical virilogical studies (morphology, chemical characterization, pathology, vectors, control measures) and current press releases on the "epidemic" in Australia. Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Hello all, Thank you to all who contributed info on Triarch. If anybody knows of other competitively priced prepared slide manufacturers I would appreciate their contact details also. Regards,
Hi Everyone, I have a JEOL 5300 SEM to which a beam controlled image acquistion system has been added. Images are acquired at 640x512 by frame averaging. It takes 1min 20sec. to collect five frames. This is about the same time it takes to do a Polaroid. Here is the problem. At magnifications above 5000x the images are fuzzy due to some kind of drift. At lower mags the drift is not noticeable. Polaroids do not show the drift. The drift rate is 100-500u per five minutes. I have the SEM manufarturer working on this too. So far I have not gotten acceptable results Now for the questions. Has anyone had this problem on other image acquisition systems? Am I trying to do something that can't be done due to drift that all SEMs have to some degree? Thanks for your answers in advance.
Gregory Rudomen Greg-at-umic.umic.sunysb.edu University Microscopy Imaging Center S.U.N.Y. at Stony Brook
Message-Id: {9602150427.AA05806-at-lisa.polymtl.ca} X-Sender: caron-at-lisa.polymtl.ca X-Mailer: Windows Eudora Version 1.4.4 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I am looking for good references whose discuss about the experimental setup (hardware, specimen preparation, experimental conditions, bias etc.) for voltage contrast and EBIC experiments. I am particularly involved in the fabrication of III-V high-speed devices for signal capture (CCD and Sample&Hold).
Mario
******************************************************* Mario Caron, M.Sc.A. ing. Research assistant Engineering Physics dept Laboratory for the Integration of Sensors and Actuators Ecole Polytechnique de Montreal C.P. 6079, succ. `centre-ville` Montreal, Quebec H3C 3A7 http://lisa.polymtl.ca
".....to form yellow, iridescent hexagonal prisms of lead iodide....."
I stained one grid with our lead stain alone, and another with our uranyl acetate stain alone. The lead stained grid had the hexagonal deposits and the other did not. So it's not the uranyl acetate. It's interesting that lead iodide forms hexagonal crystals. *******************
".....Perhaps these are your crystals - a bit of a long shot since you did not mention iodine in our elemental analysis of the crystals...."
No, there is no iodine in either of our stains, or any of the sample preparation steps; and I did not get an iodine peak with the microprobe. *******************
I have borrowed an anti static gun like the Zerostatic Eliminator that you suggest. We are in the process of trying it out. At least the grids don't jump up and cling to the lid of the plastic petri dish any more.
I'd like some input, opinions etc. on the pitfalls of having multiple operators on an SEM. My situation is that I am part of a materials analysis group that provides analytical services to corporate technology. Our group consists of a number of engineers and technicians with expertise in a variety of analytical areas. Our company has recently hired a young PhD to do materials research in another department . He is pushing very hard to be allowed to use our SEM at night and on weekends. He has 'used an SEM before' and doesn't see a problem in using ours. It has always been our practice to encourage researchers to come into the lab while their samples are being analyzed, but not to allow individuals to use the instrument themselves. I believe that the analysis is done more effectively by individuals whose expertise is in that area. However, management appears to be leaning towards changing the rules for this individual, and having me train him on the instrument.
We have a four year old JEOL 6400 instrument with a Link eXL analyzer, UTW detector, complete stage automation. We also have a computer system attached for collecting digital images, we do not use polaroid film. While this individual has used and SEM he has not used either a JEOL or LINK system before. I feel that training would be extensive. This is currently the only SEM we have. It is used approximately 12 hours per day by myself and my staff. Quite often EDS analysis is carried out using computer control and stage automation overnight. However, management feels that if someone is willing to utilize the instrument in those few hours a week when it is not being used, they need to take advantage of that to receive a maximum benefit from the asset.
Several years ago we had two older SEMS and due to 'corporate downsizing', we had only one operator. At them time I trained approximately 2 dozen engineers who desired to use the 'extra' SEM on their own. We disposed of the instrument about a year ago when it had not been touched for almost a year. The time I spent in training all of those individuals was probably greater than the time they as a group spent using the instrument. Hindsight being perfect I wish we had kept the old instrument!
Anyway, I would appreciate any input both pro and con from the list regarding opening up an instrument to multiple infrequent users.
These things help a great deal when using a glass knife. We use lecithin in the epoxy accelerator . ( use eual weight of lecithin and accelerator, then double the volume for use) A glass knife will last a long time if you have it in the resin. It was a great help to students just learning to section, They could get usable stuff the first day at the microtome. It can give a blotchy look to empty resin areas or those that have little density, but most tissues are "busy " enough that you never notice it. See Mollenhauer , 1986, J. EM Tech 3:217-222 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Dear Bonnie, I run a Materials Engineering EM lab at the University of British Columbia, with two SEMs and a TEM. Most of the researchers who use the instruments are graduate students and Research Engineers. I have always encouraged these people to do their own research on the SEMs, as much as possible, since only they know exactly what they want and that way one me can keep three instruments optimally used. After I show them how to use the SEM, I watch and encourage them to ask me for further instruction for picture taking, higher mag, etc. Some people, who have a lot of work to do, may gain my permission to use the SEM after hours, but only after I feel they have gained a lot of experience during working hours, when I am there to supervise, and only after I have instructed them on how to properly shut down and what to do in the event of problems. I also purchased the instruments originally with ease-of-use in mind. They are fully automatic and easy to get good results on. I must feel that the person has a good understanding of the important issues and knows how to properly handle the instrument. Mind you, I don't have a field emission SEM. I have had damage, but only once in 15 years and it could have happened in normal hours.. It also helps to scare them, be "the ogre of the EM lab". Mind you, we are a teaching lab, so the learning is as important as the doing. I know many EM operators do not like the idea of letting any ham-fisted graduate student or engineer loose on their instrument, but modern SEMs really can be operated by any intelligent being. So long as you satisfy yourself as to this person's knowledge, experience and caring, I'd see no harm. Hope this helps.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Hello Everyone, I'd just like to put in my two cents on the subject of multiple users of an SEM (or in my case an electron microprobe)
Our instrument is used at least 16 hours a day and since I'm only paid for eight almost all our users are trained to use the machine independantly. This allows them faster access to the machine and saves them or their advisors money. The only people we don't encourage to become independant users are those who will only use the machine once or twice. Most users require 3-4 daytime shifts before i will let them use the machine independantly (I give them my home phone number too).
In the six years I've been running the lab I can't recall one incident where the machine has actually been damaged by inexperienced users. On several occasions it has been temporarily put out of commission (computer problems, etc) but usually I can get it back on line immediately the next day. Although there is the potential for a user to damage the instrument (i.e during a sample change) most of the rest of the instrument is fairly fool proof. It is easy to screw up your analyses, but difficult to hurt the machine.
I'm sure the machine would last longer and require less service if I was the only one operating it, but we don't have that option. I also feel that users are much better off acquiring their own data, they know what they want and can change their strategy if they find something unexpected
I have prototype LVSEM that employs the stage mechanism from a Philips EM 430 TEM. I would like to purchase a cold stage for this instrument if I can find one (standard Philips rod). Anyone out there about to "decommission" one?
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
At 1:09 PM 2/9/96 -0500, Bonnie Davis - Kennametal Inc. wrote: } I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM. My situation is that I am part of a materials } analysis group that provides analytical services to corporate } technology. Our group consists of a number of engineers and technicians } with expertise in a variety of analytical areas. Our company has } recently hired a young PhD to do materials research in another department . } He is pushing very hard to be allowed to use our SEM at night and on weekends. } He has 'used } an SEM before' and doesn't see a problem in using ours. It has always } been our practice to encourage researchers to come into the lab while } their samples are being analyzed, but not to allow individuals to use the } instrument themselves. I believe that the analysis is done more effectively } by individuals whose expertise is in that area. However, management } appears to be leaning towards changing the rules for this individual, and } having me train him on the instrument. } } We have a four year old JEOL 6400 instrument with a Link eXL analyzer, } UTW detector, complete stage automation. We also have a computer system } attached for collecting digital images, we do not use polaroid film. } While this individual has used and SEM he has not used either a JEOL or } LINK system before. I feel that training would be extensive. This is } currently the only SEM we have. It } is used approximately 12 hours per day by myself and my staff. Quite } often EDS analysis is carried out using computer control and stage } automation overnight. However, management feels that if someone is } willing to utilize the instrument in those few hours a week when it is } not being used, they need to take advantage of that to receive a maximum } benefit from the asset. } } Several years ago we had two older SEMS and due to 'corporate } downsizing', we had only one operator. At them time I trained } approximately 2 dozen engineers who desired to use the 'extra' SEM on } their own. We disposed of the instrument about a year ago when it had } not been touched for almost a year. The time I spent in training } all of those individuals was probably greater than the time they as } a group spent using the instrument. Hindsight being perfect I wish we had } kept the old instrument! } } Anyway, I would appreciate any input both pro and con from the list } regarding opening up an instrument to multiple infrequent users.
Bonnie, Since we encourage use of the JEOL TEM and SEM by trained users, we run into this situation often. Currently, we require training on our instruments plus a minimum of 15 hours of daytime use prior to an approval of use during off hours. The principal investigator (of the grad. student or postdoc) is also asked to agree to cover costs should any malfunction necessitate lengthy (costly) alignment procedures. I constructed a form for this purpose and the signed agreement is kept with the log book at the scope. This has given me some control over the situation and those users who agree to our demands are usually competent users. If the SEM uses a LaB6 or is a FE, I would be more cautious, i.e., demand more daytime use and obtain written consent to replace the crystal/filament. One last thing to consider---an estimate of costs incurred by your unit during the required "daytime" use if your unit does not charge other units for instrument use. Regards, Rosemary Walsh
I tried almost every possibility on our Baltec RES010. I have found that (a), a longer time at low energy with one gun set to a low angle and one gun to a rather high angle worked best on my ceramic materials.
Andreas
______________________________________________________________ Andreas Loewe Tel: +49-228-550-355 University of Bonn Fax: +49-228-678-413 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
I'm responding to Greg Rudomen's posting about fuzzy digital images on his JEOL 5300.
Greg, can you give us some more details on your problem? It sounds like you are using a slow scan to do your frame averaging since you say it takes 1 min. 20 sec. to collect five frames.
Are you really seeing drift? Drift will generally be blurred in one direction; if the images are just not sharp, that may be another problem.
When you say the Polaroids do not show the drift, are these Polaroids of the analog or the digitized image? If the analog image is okay, something is happening during digitization.
Another item to check is the power and magnetic field situation in your lab. If the drift is more noticeable at low accelerating voltages, you may have a field problem.
Feel free to contact me directly about this. I'm using a JEOL 6300F with a Vision system, and it's no problem to obtain a clean 100K digital image.
Jim Stets Air Products and Chemicals, Inc. Allentown, PA stetsjr-at-ttown.apci.com
Ye Olde Disclaimer: I speak for myself, not my employer.
I was once employed at Baylor College of Medicine in Houston, Texas in the Division of Molecular Virology. One proffessor there did many studies on Calicivirus. Her name is Mary K. Estes. I do not know her email address but it is avaiable through Baylor's Web page address'. Hope you can obtain the info you need through her.
Best of Luck, Ed Calomeni Medical College of Ohio Dept. Pathology Toledo, OH emlab-at-opus.mco.edu
Regarding the apparent drift problem in Greg Rudomen's images:
We have seen similar effects with a JEOL 6100 interfaced to a Voyager EDS/image acquisition system. The effect is also noticeable when using the image capture features of the SEM itself. Slow scans over a minute or two, whether for a Polaroid or for digitizing, do not show the problem, as the digitization occurs only once at each beam location and a small drift would be unnoticeable in terms of seeing a distorted image; however, for elemental mapping using single scans, with long dwell times resulting in acquisitions taking hours, the drift/distortion is extremely evident. Averaging, integrating or other algorithms which entail multiple scans show the diffuseness very strongly. Most frustrating is that the problem is sometimes worse during some sessions compared to others, with no obvious differences in specimens or operating conditions.
This problem was suggested by the SEM manufacturer to be one of charging or poor grounding in our specimens. Not likely, though, as we were observing Si and GaAs crystals when the problem was most noticeable in images. The problems were most notable with these because the magnifications were considerably higher than other routine work done here. Decreasing the beam energy to ca. 1-5 keV did not improve the problem, suggesting that charging plays no role.
The problem is as yet unresolved, but I wanted to add our observations. For the time being, I acquire images using slow scans only. Maybe it has something to do with imaging on every third Tuesday of months ending with the letter "y"...
Message-Id: {199602151731.LAA05955-at-mailhub.iastate.edu} X-Sender: wes-at-pop.ameslab.gov X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I got caught with some drift the other day on an E-SEM. The image was drifting slowly in one direction. I half suspected charging, although the drift was only in one direction and varying the pressure and cutting back the beam did not help.
The problem was eventually tracked back to the modeling clay that I had used to support the irregularly shaped sample. I was not aware that it outgassed fast enough to change dimensions in the SEM. Now I know better. ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091 E-Mail: wes-at-ameslab.gov (or: wesaia-at-iastate.edu)
coal characterization and processing electron microscopy, x-ray analysis, image analysis computer applications
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Hi all, I snipped the above lines from Ya Chen's response to Bonnie. As Nestor posted last week, there have been some difficulties with the server, and it seems that I have missed some of the posts. I hold a position at the University of Iowa's Central Microscopy Research Facility. We are a multi-user resource for microscopy related techniques. We have two SEMs that we train people to operate. ANYONE can use our facility, as long as we train them, or/and they prove themselves competent in the operation of the instrument. At that point, they are issued keys, and can reserve the instrument to suit their schedule. We have never had to ask for keys back from an individual, and any problems that have occured were corrected with additional training. We do have service contracts on our microscopes, and I can be fairly handy with a tool box. Knock on wood, we have yet to have a catastrophic event (one SEM is a field emitter). With the robust design of modern electron microscopes, I don't anticipate any, either. For more on the operating philosophy of our lab, as well as instrumentation available, I suggest you visit our website at http://www.uiowa.edu/~cemrf/.
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Hi all, I snipped the above lines from Ya Chen's response to Bonnie. As Nestor posted last week, there have been some difficulties with the server, and it seems that I have missed some of the posts. I hold a position at the University of Iowa's Central Microscopy Research Facility. We are a multi-user resource for microscopy related techniques. We have two SEMs that we train people to operate. ANYONE can use our facility, as long as we train them, or/and they prove themselves competent in the operation of the instrument. At that point, they are issued keys, and can reserve the instrument to suit their schedule. We have never had to ask for keys back from an individual, and any problems that have occured were corrected with additional training. We do have service contracts on our microscopes, and I can be fairly handy with a tool box. Knock on wood, we have yet to have a catastrophic event (one SEM is a field emitter). With the robust design of modern electron microscopes, I don't anticipate any, either. For more on the operating philosophy of our lab, as well as instrumentation available, I suggest you visit our website at http://www.uiowa.edu/~cemrf/.
Alternate-Recipient: allowed Auto-Forwarded: prohibited Content-Return: allowed Disclose-Recipients: prohibited Conversion: allowed Importance: normal Priority: normal Sensitivity: Company-Confidential
POSTDOCTORAL POSITION (US citizens only)
The Physical Metallurgy Branch of the Naval Research Laboratory is currently looking for a postdoctoral researcher to study solid-solid phase transformations. The ideal candidate would have a strong background in both transmission electron microscopy and materials science (preferably with a concentration in physical metallurgy). Current research interests center on the study of solid-solid phase transformations in conventional and advanced metallic alloys, ranging from fundamental studies of martensite and precipitate morphologies in steels to microstructural analysis of low-carbon steel weldments, advanced shape memory alloys, new beta-titanium alloys, and metallic thin film and multilayer materials. Please contact either myself (Fonda-at-anvil.nrl.navy.mil, 202-767-2622) or Dr. George Spanos (Spanos-at-anvil.nrl.navy.mil, 202-767-5799) for further details about these and other research programs.
EQUIPMENT JEOL 200CX, Philips CM30, and Hitachi 9000 TEMs Isothermal heat treatment facility for the study of rapid phase transformations in an inert environment Quenching and deformation dilatometer Hitachi FEG-SEM Auger electron spectroscope
PROGRAM Descriptions of the postdoctoral programs at NRL may be obtained from Lesley Renfro, Program coordinator, Code 1005.7, Naval Research Laboratory, Washington, DC, 20375 (Renfro-at-utopia.nrl.navy.mil, 202-767-3865). Dr. Spanos is listed as research advisor for the NRC program descriptions.
ELIGIBILITY The postdoctoral programs at NRL are open only to US citizens.
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
Could someone explain the difference between DEPEX and DPX mounting media or are they used interchangeably? If you have a fluorescently labelled sample cleared in methyl salicylate, does anyone have a protocol for mounting in DPX? Do you have go back to xylene before mounting in DPX or can you go directly to DPX from methyl salicylate? How long does the DPX taken to harden on a 100 um sample? Thanks.
Patty Jansma Tel:602-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
The following meeting may be of interest to some readers of this listerv.
WORKSHOP
PCR-IN SITU HYBRIDIZATION: =20 What you need to know to make it work
As part of the Histochemical Society Annual Meeting,=20 August 2-3, 1996,=20 Place: Hyatt Regency Hotel, Bethesda, MD
=09Topic: The use of PCR has revolutionized study of low-abundance=20 DNA and RNA molecules and recent advances have made it possible to=20 amplify DNA and cDNA targets in fixed cells and paraffin-embedded tissues= =20 on slides. The use of these techniques is beginning to have a major=20 impact in both cell biology and diagnostic pathology. The workshop=20 will deal with: =20 =09=A5methodologies to label and detect nucleic acids using=20 non-radioactive primers=20 =09=A5advantages and disadvantages of PCR-ISH, pitfalls, controls,=20 optimal protocols, effects of fixatives, protease digestion, primers,=20 nucleotides, relevant enzymes and related issues. =20 =09=A5planned format includes lectures, question periods and=20 interactive demonstrations of in situ-PCR, PCR-in situ hybridization=20 and reverse transcriptase-in situ PCR. Thermocycling systems appropriate= =20 for processing selected tissue specimens will be utilized and will=20 generally be available during the workshop. =20 =09=A5A syllabus will be provided for workshop participants.=20
=09Organizer: G. J. Nuovo, SUNY at Stony Brook
A related symposium will be on=20
ENHANCEMENT METHODS FOR IN SITU HYBRIDIZATION=20
=09Topic: Fluorescent in situ hybridization (FISH) techniques have=20 been utilized for the past decade and offer a widely used microscopical= =20 tool providing high sensitivity and resolution, as well as the ability to= =20 detect multiple targets. To increase the sensitivity of FISH, indirect=20 amplification by immunological methods has often been the method of=20 choice. This symposium will include the presentation of several novel=20 approaches using the enzymatic deposition of fluorescent reporter=20 molecules to amplify target localization. These methods may offer a new=20 approach to increase both the detection, resolution, and sensitivity of=20 the FISH method.=20 =09Organizer: S.L. Erlandsen =09Speakers:=20 =09S. L. Erlandsen, University of Minnesota, Introduction. =09A.K. Raap, Leiden University, Sensitive and high resolution FISH=20 =09B. Schmidt, Carnegie Mellon University, Signal amplification in=20 the detection of single copy DNA + RNA by enzyme catalyzed deposition=20 (CARD) of the novel fluorescent reporter substrate, Cy 3.29-tyramide. =09V.L. Singer, Molecular Probes, Inc., The ELF alkaline=20 phosphatase substrate provides a bright, photostable, fluorescent signal=20 amplification method for FISH.=20
Another symposium will be on=20
APPLICATIONS OF MOLECULAR TECHNOLOGIES TO DIAGNOSTIC AND INVESTIGATIVE=20 PROBLEMS OF BREAST CANCER.=20
=09Topic: The symposium will include presentations on in situ=20 analysis of control of cell proliferation in breast cancer, molecular=20 markers of apoptosis and their correlation with hormone receptor=20 expression and other histologic prognostic markers and the structure of=20 breast tissue as it related to breast cancer development.=20 =09Organizer: A. M. Gown =09Speakers:=20 =09H. Wolfe, Tufts University, Molecular markers of apoptosis and=20 their correlation with hormone receptor expression and other histologic=20 prognostic markers. =09R. Jensen, Vanderbilt University, BRCA1 gene expression in breast cancer cells. =09A.M. Gown, University of Washington, In situ analysis of control=20 of cell proliferation in breast cancer. =09J.W. Gray, UC San Francisco, Applications of molecular techniques=20 to diagnostic and investigative problems of breast cancer.
POSTERS ON TOPICS RELATED TO THE WORKSHOP AND SYMPOSIA are welcome =20 (abstract deadline: March 15, 1996) and abstracts will be published as=20 Proceedings of the Histochemical Society Meeting in the Journal of=20 Histochemistry and Cytochemistry. .
CONTACT:=20 Inquiries about program matters and requests for registration materials=20 or abstract forms, and hotel accommodations should be sent to: The=20 Histochemical Society, 4 Barlows Landing Rd., Pocasset, MA 02559=20 (Telephone: 508-563-1155; FAX 508-563-1211 or e-mail: lmaser-at-mbl.edu). =20 or inquiries related to the scientific program to: Dr. W.L. Stahl=20 (e-mail: wlstahl-at-u.washington.edu).=20 =20
Dear Greg, Can you see the image drift at high mag. in TV-rate? Is this perhaps EM field ripple? or mechanical vibration? The image acquisition system can only acquire what the SEM puts out, so this drift will certainly wipe out the benefits of frame averaging. If it takes so long to acquire a frame-averaged image, why not just acquire one long frame? I have a passive system and usually take a 10 sec., 1024X768 single frame, but the image certainly shows up any drift of the SEM stage. This can always be traced to the specimen not being secured well enough. Sticky tabs are often guilty of this. If the SEM doesn't drift, the image is perfect to 50,000X.
One problem I have had in the past is bad vibration problems at 20 minutes past the hour on class days. This is when classes change and two hundred undergrad engineers tromp through the halls. People doing critical hi-res, low kV work know to do it after hours, when the building is quieter and the elevators aren't running. Good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
I am looking for a mounting medium which can be used for coverslipping Giemsa stained cells containing latex microspheres. Xylene containing mounting media cannot been used because of the solubility of the latex microspheres. On the other hand, water soluble mounting media, like gelatin/glycerin, destain the Giemsa. The staining is important to me to clearly locate (for counting) the microspheres within the cell borders. Counting without coverslip can be done but I prefere using a 40x/1.0 objective (oil).
I'm looking for other microscope pools, where teaching microscopes are shared to maximize usage. I've worked for 20 years at Microscope Services at the University of California, Davis. We have about 2100 optical microscopes of high quality, and move 600-700 around every quarter from classrooms where they aren't needed to those where they are. We also provide preventive maintenance and repairs, short courses microscopes for research, recharging for our services. This arrangement works well at UC Davis, so it must be duplicated at other schools. I just don't know where. If any of you know of schools with similar arrangements, would you please post a contact for me? I would very much like to hear from someone with similar tasks and problems. Richard L. Markgraf Microscope Services University of California, Davis Ph. (916)752-3477 Fax (916)752-6363 rlmarkgraf-at-ucdavis.edu
Joint Annual Meeting of the Microscopy Society of America, Microbeam Analysis Society and the Microscopical Society of Canada/Societe de Microscopie du Canada, August 11-15, 1996, Minneapolis, Minnesota.
Deadline for receipt of Extended Abstracts: March 15, 1996.
Approximately 7,000 Registration Bulletin / Call for Papers packages have been mailed, most on February 2, to: the memberships of the three Sponsoring Societies; the memberships of four Local Affiliates of the Microscopy Society of America (Northwestern Ohio Microscopy Society, Minnesota Microscopy Society, Southern California Society for Electron Microscopy - Technologists, and the Iowa Microscopy Society); individuals who attended Microscopy & Microanalysis '95, Kansas City; and other individuals who have requested one.
Please contact our office if you should have received the Registration Bulletin / Call for Papers but haven't, or if you would like to. We'll respond to your request immediately. Please note that the deadline for receipt of Extended Abstracts is March 15, 1996.
Hope to see you in Minneapolis,
Larry Maser
Microscopy & Microanalysis '96 4 Barlows Landing Rd., Suite 8 Pocasset MA 02559
Message-Id: {199602161628.KAA13875-at-mailhub.iastate.edu} X-Sender: wes-at-pop.ameslab.gov X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
From another university lab:
We too allow users on the scope after training as many in academic settings appear to do. I routinely query the potential users about the realistic duration of their work as we have trained some at their insistence only to find out they have no specific project in mind, or that their actual SEM usage was not nearly what they first imagined. Of course, since they are charged for operator assistance, I guess we will continue to train them if they care to pay for it.
Now a question, We have adopted the practice of leaving the filament current control on our JEOL 840A at the proper saturation point (for our normal 15 kv operating point) and just turning off the high voltage control for exchanging samples. We have thus avoided problems from inexperienced operators over-saturating. It appears to be working as our filament lifetimes have increased to the values for when only experienced users operated the scope. My question to the list is whether there might be any adverse consequences from this practice as there do not appear to be any.
Immediate Opening for SEM Analysis Engineer at SVC:
Principal Duties & Responsibilities:
SEM analysis of display components; basic failure analysis. Responsible for upkeep of JSM 820, with full JEOL service contract; Supervision & training of other SEM users.
Special work circumstances:
It is preferred that candidate be able to work either an early (start -at- 6AM) or late (finish -at- 8PM) shift.
Experience & Education Requirements:
B.S. in Physical Science or Engineering or equivalent. Minimum 3 yrs. experience in Electron Microscopy.
Company Background:
In just 4 years Silicon Video Corp. has become on of the hottest start ups in Silicon Valley. We are developing a new class of flat panel displays - Thin CRT. Millions in funding from the government have been obtianed. Large corporate partners in industry and academia connections have been secured. We're searching for highly qualified candidates to join our team.
Hello All, A graduate student here has been given the task of looking at the surface of a highly polished stainless steel ball which is used in hip replacements. The ball is about 2.5-3 cm in diameter and as you might imagine, is extremely reflective. Since all I have ever examined in an SEM are biological samples, I haven't been able to help her very much. This morning we got what appeared to be a grid image on the surface and I am wondering if this might not be a reflection of the interior surface of the specimen chamber. I suggested sputter coating the surface to cut down on reflection and mounting it on some double stick tape to keep it stable; but she doesn't think the person who assigned her this task would want that. Without regard to the wants of the person assigning the task, do any of you in the materials science have any suggestions which might be helpful to this young lady. She and I would both be very grateful. TIA, Sandra Zane Sandra F. Zane, EM Tech. sfzane-at-unccvm.uncc.edu Dept of Biol., UNC Charlotte (704) 547-4051 9201 University City Blvd. Fax (704) 547-3128 Charlotte, NC 28223-0001
} Hi Everyone, } I have a JEOL 5300 SEM to which a beam controlled image } acquisition system has been added. Images are acquired at } 640x512 by frame averaging. It takes 1min 20sec. to } collect five frames. This is about the same time it takes } to do a Polaroid. } Here is the problem. At magnifications above 5000x the } images are fuzzy due to some kind of drift. At lower mags } the drift is not noticeable. Polaroids do not show the } drift. The drift rate is 100-500u per five minutes. I } have the SEM manufarturer working on this too. So far I } have not gotten acceptable results } Now for the questions. Has anyone had this problem on } other image acquisition systems? Am I trying to do } something that can't be done due to drift that all SEMs } have to some degree? } Thanks for your answers in advance. } } Gregory Rudomen } Greg-at-umic.umic.sunysb.edu } University Microscopy Imaging Center } S.U.N.Y. at Stony Brook }
Gregory, There are 3 kinds of averaging techniques used in SEM digitization: frame averaging, line averaging and pixel averaging. For most of "built-in" acquisition system, the frame capture is used, so only support frame averaging. The drawback for using frame averaging is to produce a fuzzy image as long as drafting exist. Pixel averaging is mostly suitable to overcome this situation.
Ya Chen
Ya Chen Cryo/SEM project Coordinator TEL : 608-263-8481 Integrated Microscopy Resource (IMR) FAX : 608-265-4076 University of Wisconsin-Madison Email:YChen-at-macc.wisc.edu IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr/imr.html IMR Symposium on integrated microscopy '96: Sept. 20-22, 1996
Dear Sandra, It is very difficult to look at the surface of any highly-polished metal, since there is nothing to focus on. However, you cannot get a reflection image in an SEM except by charge buildup, so if you did get a grid image from your secondary detector, it implies the ball is not well grounded. If you stick it down, use conductive sticky-tabs or make a connection useing silver or carbon dag. One help to focusing is to draw on the top surface with a felt-tip pen, so you can find the initial focus. My experience though, is that a good reflextive surface gives you very little to see, even at high mag. Good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
} Now a question, } We have adopted the practice of leaving the filament current control on our } JEOL 840A at the proper saturation point (for our normal 15 kv operating } point) and just turning off the high voltage control for exchanging samples. } We have thus avoided problems from inexperienced operators over-saturating. } It appears to be working as our filament lifetimes have increased to the } values for when only experienced users operated the scope. My question to } the list is whether there might be any adverse consequences from this } practice as there do not appear to be any. Dear Warren, We always leave the filament current up to the saturation point (or just under it for maximum filament life). In fact, to stop little fingers, I removed the filament current knob from my console years ago. I can change it with a screwdriver when I need to. I always turn the acc. voltage on and off and I get at least a month out iof a filament at 20kv. I don't know if I would do it on my 200kv TEM, but SEM is fine. Best of luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Contents Retrieved from Microscopy Listserver Archives